IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. This is a Cascade Kinase assay, however no details of the method(s) used are given here, nor in quoted/associated references. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-patu Human-PATU Pancreatic Adenocarcinoma cell line patu PATU-Human Pancreatic adenocarcinoma cell line anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. There are several human pancreatic adenocarcinoma cell lines with "PaTu" in the name; it is not stated which one is used in Figure 1A; Supplementary Figure 1A uses Patu 898T . The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay sequence tag sequence tag identification sequence tag Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. Human BRAF alone, and with MEK proteins were incubated with and without ATP, separated by SDS-PAGE and samples from the gel analyzed by LC-MS/MS. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-hpac HPAC Human pancreatic adenocarcinoma cell line hpac HPAC Human pancreatic adenocarcinoma cell line http://www.lgcstandards-atcc.org/products/all/CRL-2119.aspx?geo_country=gb ATCC CRL-2119 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. There are several human pancreatic adenocarcinoma cell lines with "PaTu" in the name; it is not stated which one is used here. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro fps fluorescence polarization spectroscopy Fluorescence anisotropy FPS FPS Fluorescence anisotropy fps predetermined predetermined participant predetermined Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-a375 Human malignant melanoma a375 Human malignant melanoma ATCC catalog # CRL-1872 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-panc1 Homo sapiens pancreatic carcinoma cell line PANC-1 panc1 Human pancreatic carcinoma cell line PANC-1 Established from a pancreatic carcinoma of ductal origin from a 56-year-old Caucasian male. Cells possess the type B phenotype for G6PD. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. There are several human pancreatic adenocarcinoma cell lines with "PaTu" in the name; it is not stated which one is used here. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-hey Homo sapiens hey hey anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. The cell line stated to have been used here -"Hey1" has been entered as "HEY", as no cell line "hey1" or "Hey-1" can be found. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. No experimental protocol is given for this interaction; Participant detection has been assumed to be Western Blotting. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-nci_h1666 Human Human NCI-H1666 bronchoalveolar adenocarcinoma nci_h1666 Human NCI-H1666 bronchoalveolar adenocarcinoma http://www.lgcstandards-atcc.org/products/all/CRL-5885.aspx?geo_country=gb ATCC = CRL-5885 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-colo_357 Human lymph node metastasis adenocarcinoma pancreas colo_357 Human lymph node metastasis adenocarcinoma pancreas anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. There are several human pancreatic adenocarcinoma cell lines with "PaTu" in the name; it is not stated which one is used here. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-hct116 Human colon carcinoma HCT116 cells hct116 Human colon carcinoma line HCT116 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-a_549 Homo sapiens Caucasian lung carcinoma cell line a_549 Human Caucasian lung carcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. There are several human pancreatic adenocarcinoma cell lines with "PaTu" in the name; it is not stated which one is used here. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-mda_mb_231 Homo sapiens Caucasian breast adenocarcinoma mda_mb_231 Human Caucasian breast adenocarcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-hs294t Human HS294T-Melanoma cell line from metastatic site: lymph node hs294t Human HS294T-Melanoma cell line from metastatic site: lymph node http://www.lgcstandards-atcc.org/products/all/HTB-140.aspx?geo_country=gb ATCC- HTB-140 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro fluorescence fluorescence technology fluorescence predetermined predetermined participant predetermined Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The method is stated to be homogeneous time-resolved fluorescence (hTRF)". Figure 6 legend states that ATP was present in all conditions except for the no ATP control. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight by comassie molecular weight estimation by coomasie staining weight by comassie Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro kinase htrf kinase homogeneous time resolved fluorescence kinase HTRF homogeneous time-resolved fluorescence kinase htrf predetermined predetermined participant predetermined Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. Phosphorylation of MEK was measured by TR-FRET using CisBio reagents;an antibody to pMEK is labelled with a Europium containing compound (fluorescence Donor) and an anti-GST antibody labelled with "d2" - an organic moiety - as the fluorescence acceptor. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-hec1a Homo sapiens hec1a human endometrial carcinoma cells anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. Cancer Cell (1535-6108) 2014 imex curation Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions human-mia_capa Human MIA-CaPa Pancreatic Carcinoma cell line mia_capa MIA PaCa-2 Pancreatic carcinoma cell line http://www.lgcstandards-atcc.org/products/all/CRL-1420.aspx?geo_country=gb ATCC CRL-1420 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Haling J.R., Sudhamsu J., Yen I., Sideris S., Sandoval W., Phung W., Bravo B.J., Giannetti A.M., Peck A., Masselot A., Morales T., Smith D., Brandhuber B.J., Hymowitz S.G., Malek S. Several cell lines used are stated to have mutations- BRAF V600E and P-loop; however it is not known if these cell lines are homozygous or heterozygous for these mutations, so mutations have not been entered here, as it is not known if the WT or mutant is interacting, or both. Results from Supplementary Figure 4B (showing Kinase activity of recombinant wild-type BRAF and BRAFV600E kinase domain +/- dimer defective mutant R509H) have not been entered here as no experimental details are given, such as the substrate used, inter al.. Figure 1/S1 - I have no idea which cell lines are meant to show positive results - so have entered "best guesses". In some IPs there are +ve results, but NOT in other apparently identical IPs. FIGURE SUPP 5B IS IN VITRO - text says somewhere in main body of the paper. Email sent 27/10 - will be amazed if I get a reply...( The only reply was -quote "all were human") ...far too many Qs asked)b Sorry - this is probably rather a mess.. The authors have confirmed that all constructs used are of Human origin. It is not known for experiments involving Mass Spectrometry and SPR, whether full length or n-terminally deleted MEK1 was used. hymowitz.sarah@gene.com,malek.shiva@gene.com Accepted 2014-DEC-04 by ORCHARD Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation gdc-0879 GDC-0879 small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) raf1_human RAF proto-oncogene serine/threonine-protein kinase RAF1 RAF Proto-oncogene c-RAF Raf-1 protein protein human Homo sapiens Human MEHIQGAWKTISNGFGFKDAVFDGSSCISPTIVQQFGYQRRASDDGKLTDPSKTSNTIRVFLPNKQRTVVNVRNGMSLHDCLMKALKVRGLQPECCAVFRLLHEHKGKKARLDWNTDAASLIGEELQVDFLDHVPLTTHNFARKTFLKLAFCDICQKFLLNGFRCQTCGYKFHEHCSTKVPTMCVDWSNIRQLLLFPNSTIGDSGVPALPSLTMRRMRESVSRMPVSSQHRYSTPHAFTFNTSSPSSEGSLSQRQRSTSTPNVHMVSTTLPVDSRMIEDAIRSHSESASPSALSSSPNNLSPTGWSQPKTPVPAQRERAPVSGTQEKNKIRPRGQRDSSYYWEIEASEVMLSTRIGSGSFGTVYKGKWHGDVAVKILKVVDPTPEQFQAFRNEVAVLRKTRHVNILLFMGYMTKDNLAIVTQWCEGSSLYKHLHVQETKFQMFQLIDIARQTAQGMDYLHAKNIIHRDMKSNNIFLHEGLTVKIGDFGLATVKSRWSGSQQVEQPTGSVLWMAPEVIRMQDNNPFSFQSDVYSYGIVLYELMTGELPYSHINNRDQIIFMVGRGYASPDLSKLYKNCPKAMKRLVADCVKKVKEERPLFPQILSSIELLQHSLPKINRSASEPSLHRAAHTEDINACTLTTSPRLPVF DIP protein P04049 original sequence version: 117 DIP protein P04049 original sequence version: 106 DIP protein P04049 original sequence version: 114 DIP protein P04049 original sequence version: 141 DIP protein P04049 original sequence version: 142 DIP protein P04049 original sequence version: 147 DIP protein P04049-1 original sequence version: 136 DIP protein P04049 original sequence version: 136 DIP protein P04049 original sequence version: 153 DIP protein P04049 original sequence version: 150 DIP protein P04049 original sequence version: 164 DIP protein P04049 original sequence version: 172 DIP protein P04049-1 original sequence version: 172 DIP protein P04049 original sequence version: 184 DIP protein P04049 original sequence version: 188 DIP protein P04049 original sequence version: 195 EF821B5349711BC3 mp2k1_human Dual specificity mitogen-activated protein kinase kinase 1 ERK activator kinase 1 MAPK/ERK kinase 1 MAP2K1 MEK1 PRKMK1 protein protein human Homo sapiens Human MPKKKPTPIQLNPAPDGSAVNGTSSAETNLEALQKKLEELELDEQQRKRLEAFLTQKQKVGELKDDDFEKISELGAGNGGVVFKVSHKPSGLVMARKLIHLEIKPAIRNQIIRELQVLHECNSPYIVGFYGAFYSDGEISICMEHMDGGSLDQVLKKAGRIPEQILGKVSIAVIKGLTYLREKHKIMHRDVKPSNILVNSRGEIKLCDFGVSGQLIDSMANSFVGTRSYMSPERLQGTHYSVQSDIWSMGLSLVEMAVGRYPIPPPDAKELELMFGCQVEGDAAETPPRPRTPGRPLSSYGMDSRPPMAIFELLDYIVNEPPPKLPSGVFSLEFQDFVNKCLIKNPAERADLKQLMVHAFIKRSDAEEVDFAGWLCSTIGLNQPSTPTHAAGV DIP protein Q02750 original sequence version: 96 DIP protein Q02750 original sequence version: 93 DIP protein Q02750-1 original sequence version: 151 DIP protein Q02750 original sequence version: 151 DIP protein Q02750-1 original sequence version: 166 0344118FFC842D51 atp ATP Adenosine 5'-triphosphate Adenosine triphosphate ADENOSINE-5'-TRIPHOSPHATE Adephos Adetol Adynol Atipi ATP Atriphos Cardenosine Fosfobion Glucobasin H4atp Myotriphos Triadenyl Triphosphaden adenosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) braf_human Serine/threonine-protein kinase B-raf BRAF BRAF1 RAFB1 v-Raf murine sarcoma viral oncogene homolog B1 p94 Proto-oncogene B-Raf protein protein human Homo sapiens Human MAALSGGGGGGAEPGQALFNGDMEPEAGAGAGAAASSAADPAIPEEVWNIKQMIKLTQEHIEALLDKFGGEHNPPSIYLEAYEEYTSKLDALQQREQQLLESLGNGTDFSVSSSASMDTVTSSSSSSLSVLPSSLSVFQNPTDVARSNPKSPQKPIVRVFLPNKQRTVVPARCGVTVRDSLKKALMMRGLIPECCAVYRIQDGEKKPIGWDTDISWLTGEELHVEVLENVPLTTHNFVRKTFFTLAFCDFCRKLLFQGFRCQTCGYKFHQRCSTEVPLMCVNYDQLDLLFVSKFFEHHPIPQEEASLAETALTSGSSPSAPASDSIGPQILTSPSPSKSIPIPQPFRPADEDHRNQFGQRDRSSSAPNVHINTIEPVNIDDLIRDQGFRGDGGSTTGLSATPPASLPGSLTNVKALQKSPGPQRERKSSSSSEDRNRMKTLGRRDSSDDWEIPDGQITVGQRIGSGSFGTVYKGKWHGDVAVKMLNVTAPTPQQLQAFKNEVGVLRKTRHVNILLFMGYSTKPQLAIVTQWCEGSSLYHHLHIIETKFEMIKLIDIARQTAQGMDYLHAKSIIHRDLKSNNIFLHEDLTVKIGDFGLATVKSRWSGSHQFEQLSGSILWMAPEVIRMQDKNPYSFQSDVYAFGIVLYELMTGQLPYSNINNRDQIIFMVGRGYLSPDLSKVRSNCPKAMKRLMAECLKKKRDERPLFPQILASIELLARSLPKIHRSASEPSLNRAGFQTEDFSLYACASPKTPIQAGGYGAFPVH DIP protein P15056 original sequence version: 182 DIP protein P15056 original sequence version: 133 DIP protein P15056 original sequence version: 147 DIP protein P15056 original sequence version: 173 DIP protein P15056 original sequence version: 178 DIP protein P15056 original sequence version: 188 DIP protein P15056 original sequence version: 193 DIP protein P15056 original sequence version: 195 DIP protein P15056 original sequence version: 146 DIP protein P15056 original sequence version: 153 0798C2AAB487E813 adenosine 5'-[beta,gamma-methylene]triphosphate adenosine 5'-[beta,gamma-methylene]triphosphate PHOSPHOMETHYLPHOSPHONIC ACID ADENYLATE ESTER 5'-O-(hydroxy{[hydroxy(phosphonomethyl)phosphoryl]oxy}phosphoryl)adenosine beta,gamma-Methylene ATP ADENOSINE 5'-[BETA,GAMMA-METHYLENE]TRIPHOSPHATE adenosine 5'-beta,gamma-mu-methylenetriphosphate AMP-PCP small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) UFZTZBNSLXELAL-IOSLPCCCSA-N UFZTZBNSLXELAL-IOSLPCCCSA-N g-573 G-573 small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) map2k1-braf 13 32 unspecified role unspecified role prey prey 29 unspecified role unspecified role prey prey 30 unspecified role unspecified role bait bait association association Supp Fig 1C. When all MEK1 and MEK1-BRAF complexes are immunodepleted from cell lysates, free BRAF and CRAF remain in the flow through. map2k1-braf-1 2 32 unspecified role unspecified role prey prey 30 unspecified role unspecified role bait bait 29 unspecified role unspecified role prey prey association association 1A, supp 1A. This cell line expresses a mutated form of K-Ras. Coimmunoprecipitation (coIP) of MEK1 with BRAF and CRAF revealed that endogenous BRAF-MEK1 complexes are enriched in multiple KRAS-MT and BRAF-WT/KRAS-WT cell lines, but are significantly less abundant in the BRAF-V600E and BRAF-P loop cell lines. A barely visible band is present for CRAF in Supplementary figure 1A. map2k1-braf-1 27 29 unspecified role unspecified role prey prey 30 unspecified role unspecified role bait bait 32 unspecified role unspecified role prey prey association association 1A This cell line only expressed mutated K-Ras (KRAS-MT). Coimmunoprecipitation (coIP) of MEK1 with BRAF and CRAF revealed that endogenous BRAF-MEK1 complexes are enriched in multiple KRAS-MT and BRAF-WT/KRAS-WT cell lines, but are significantly less abundant in the BRAF-V600E and BRAF-P loop cell lines. map2k1-braf-1 16 30 unspecified role unspecified role bait bait 29 unspecified role unspecified role prey prey 32 unspecified role unspecified role prey prey association association 1A This cell line only expressed mutated K-Ras (KRAS-MT). Coimmunoprecipitation (coIP) of MEK1 with BRAF and CRAF revealed that endogenous BRAF-MEK1 complexes are enriched in multiple KRAS-MT and BRAF-WT/KRAS-WT cell lines, but are significantly less abundant in the BRAF-V600E and BRAF-P loop cell lines. map2k1-braf-2 7 32 unspecified role unspecified role prey prey 29 unspecified role unspecified role prey prey 30 unspecified role unspecified role bait bait association association 1A/C, Supp fig 1A. Coimmunoprecipitation (coIP) of MEK1 with BRAF and CRAF revealed that endogenous BRAF-MEK1 complexes are enriched in multiple KRAS-MT and BRAF-WT/KRAS-WT cell lines, but are significantly less abundant in the BRAF-V600E and BRAF-P loop cell lines. Only a weak band is seen for CRAF. map2k1-braf-3 21 32 unspecified role unspecified role prey prey 29 unspecified role unspecified role prey prey 30 unspecified role unspecified role bait bait association association 1A Coimmunoprecipitation (coIP) of MEK1 with BRAF and CRAF revealed that endogenous BRAF-MEK1 complexes are enriched in multiple KRAS-MT and BRAF-WT/KRAS-WT cell lines, but are significantly less abundant in the BRAF-V600E and BRAF-P loop cell lines. map2k1-braf-4 6 29 unspecified role unspecified role prey prey 30 unspecified role unspecified role bait bait 32 unspecified role unspecified role prey prey P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain V E Wed May 02 19:57:56 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 18:27:39 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' association association 1A/C, 6D, Supp fig 1A. Coimmunoprecipitation (coIP) of MEK1 with BRAF and CRAF revealed that endogenous BRAF-MEK1 complexes are enriched in multiple KRAS-MT and BRAF-WT/KRAS-WT cell lines, but are significantly less abundant in the BRAF-V600E and BRAF-P loop cell lines. Vemurafenib - BRAF inhibitor effective only against cells with the V600E or V600 BRAF mutation. GDC-0879 -a selective B-Raf inhibitor Figure 1C; stronger bands are seen in the Cytosolic IP; otherwise CRAF bands are often weak. map2k1-braf-5 9 29 unspecified role unspecified role prey prey 32 unspecified role unspecified role prey prey P15056:p.Gly464Glu mutation mutation certain certain sequence position certain certain certain sequence position certain G E Wed May 02 14:48:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly464glu' Tue Jun 30 19:18:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly464Glu' 30 unspecified role unspecified role bait bait association association 1A Coimmunoprecipitation (coIP) of MEK1 with BRAF and CRAF revealed that endogenous BRAF-MEK1 complexes are enriched in multiple KRAS-MT and BRAF-WT/KRAS-WT cell lines, but are significantly less abundant in the BRAF-V600E and BRAF-P loop cell lines. map2k1-braf-6 15 32 unspecified role unspecified role prey prey P15056:p.Gly464Val mutation mutation certain certain sequence position certain certain certain sequence position certain G V Wed May 02 17:44:40 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly464val' Tue Jun 30 17:29:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly464Val' 30 unspecified role unspecified role bait bait 29 unspecified role unspecified role prey prey association association 1A, 6E, Supp 5C. Coimmunoprecipitation (coIP) of MEK1 with BRAF and CRAF revealed that endogenous BRAF-MEK1 complexes are enriched in multiple KRAS-MT and BRAF-WT/KRAS-WT cell lines, but are significantly less abundant in the BRAF-V600E and BRAF-P loop cell lines. GDC-0879 -a selective B-Raf inhibitor raf1-braf-15 9 29 unspecified role unspecified role bait bait possible_phospho_ser338 opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain 30 unspecified role unspecified role prey prey 32 unspecified role unspecified role prey prey P15056:p.Gly464Glu mutation mutation certain certain sequence position certain certain certain sequence position certain G E Wed May 02 15:42:04 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly464glu' Tue Jun 30 17:09:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly464Glu' association association 1B raf1-braf-16 15 32 unspecified role unspecified role prey prey P15056:p.Gly464Val mutation mutation certain certain sequence position certain certain certain sequence position certain G V Wed May 02 16:01:01 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly464val' Tue Jun 30 16:51:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly464Val' 29 unspecified role unspecified role bait bait possible_phospho_ser338 opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain 30 unspecified role unspecified role prey prey association association 1B raf1-braf-17 11 30 unspecified role unspecified role prey prey 32 unspecified role unspecified role prey prey 29 unspecified role unspecified role bait bait possible_phospho_ser338 opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain association association 1B, 6E. GDC-0879 -a selective B-Raf inhibitor braf-map2k1-3 7 30 unspecified role unspecified role prey prey 32 unspecified role unspecified role bait bait association association 1D (EGF) stimulation of HEK293 BRAF-WT cells initially results in disruption of the BRAF-MEK1 complex but this is followed by re-emergence of the BRAF-MEK complex later on. adenosine_5___beta_gamma_methylene_triphosphate-braf-1 19 34 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 33 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 30 unspecified role unspecified role neutral component neutral component c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 32 unspecified role unspecified role neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 102 99 direct interaction direct interaction Table S1, (figure 2) 2.8A working 18.64%, Free: 24.02% braf-gdc_0879 19 28 unspecified role unspecified role neutral component neutral component 32 unspecified role unspecified role neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain V E Wed May 02 15:32:17 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 18:49:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. direct interaction direct interaction Table S1 (fig 2) 2.8A working:21.46%, Free: 27.45%. braf-map2k1-4 26 32 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P15056:p.Arg462Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R E Wed May 02 14:58:58 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg462glu' Tue Jun 30 18:05:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg462Glu' P15056:p.[Arg462Glu;Ile617Arg] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R E certain certain sequence position certain certain certain sequence position certain I R Wed May 02 16:57:51 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg462glu,ile617arg' Tue Jun 30 18:04:44 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg462Glu;Ile617Arg]' P15056:p.Ile617Arg mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain I R Wed May 02 15:00:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile617arg' Tue Jun 30 17:30:46 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile617Arg' 30 unspecified role unspecified role prey prey physical association physical association 3B braf-map2k1-5 17 30 unspecified role unspecified role neutral component neutral component kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range Q02750:p.[Ser228Glu;Met230Ala] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain S E certain certain sequence position certain certain certain sequence position certain M A Wed May 02 14:52:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser228glu,met230ala' Tue Jun 30 19:51:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser228Glu;Met230Ala]' Q02750:p.Ile310Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain I E Wed May 02 18:45:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile310glu' Tue Jun 30 18:26:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile310Glu' Q02750:p.Leu314Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain L E Wed May 02 19:38:32 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu314glu' Tue Jun 30 15:46:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu314Glu' Q02750:p.Ser228Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain S E Wed May 02 17:09:36 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser228glu' Tue Jun 30 17:56:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser228Glu' Q02750:p.[Ile310Glu;Leu314Glu] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain I E certain certain sequence position certain certain certain sequence position certain L E Wed May 02 14:33:19 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile310glu,leu314glu' Tue Jun 30 15:58:10 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ile310Glu;Leu314Glu]' Q02750:p.[Ser228Glu;Met230Ala;Ile310Glu;Leu314Glu] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain S E certain certain sequence position certain certain certain sequence position certain M A certain certain sequence position certain certain certain sequence position certain I E certain certain sequence position certain certain certain sequence position certain L E Wed May 02 15:28:05 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser228glu,met230ala,ile310glu,leu314glu' Tue Jun 30 15:57:33 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser228Glu;Met230Ala;Ile310Glu;Leu314Glu]' ecoli Escherichia coli (strain K12) 32 unspecified role unspecified role neutral component neutral component kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 118 127 direct interaction direct interaction 3C, 6B, Supp 2E G-573 -a MEK inhibitor, GDC-0879 -a BRAF inhibitor, PLX-4720-a BRAF inhibitor. ATP braf-map2k1-6 1 30 enzyme target enzyme target substrate substrate neutral component neutral component c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q02750:p.Ile310Glu mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain I E Wed May 02 15:41:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile310glu' Tue Jun 30 17:21:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile310Glu' Q02750:p.Ser228Glu mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain S E Wed May 02 14:31:51 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser228glu' Tue Jun 30 19:08:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser228Glu' Q02750:p.[Ser228Glu;Met230Ala] mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain S E certain certain sequence position certain certain certain sequence position certain M A Wed May 02 19:10:13 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser228glu,met230ala' Tue Jun 30 19:46:10 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser228Glu;Met230Ala]' Q02750:p.[Ser228Glu;Met230Ala;Ile310Glu;Leu314Glu] mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain S E certain certain sequence position certain certain certain sequence position certain M A certain certain sequence position certain certain certain sequence position certain I E certain certain sequence position certain certain certain sequence position certain L E Wed May 02 17:01:47 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser228glu,met230ala,ile310glu,leu314glu' Tue Jun 30 19:14:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser228Glu;Met230Ala;Ile310Glu;Leu314Glu]' Q02750:p.[Ile310Glu;Leu314Glu] mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain I E certain certain sequence position certain certain certain sequence position certain L E Wed May 02 16:50:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile310glu,leu314glu' Tue Jun 30 17:15:38 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ile310Glu;Leu314Glu]' Q02750:p.Leu314Glu mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain L E Wed May 02 18:16:32 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu314glu' Tue Jun 30 18:16:19 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu314Glu' ecoli Escherichia coli (strain K12) 32 enzyme enzyme neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 132 141 phosphorylation phosphorylation reaction phosphorylation 3Di P15056 increases activity of Q02750 braf-map2k1-7 1 30 enzyme target enzyme target substrate substrate neutral component neutral component n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) 32 enzyme enzyme neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. phosphorylation phosphorylation reaction phosphorylation 3Dii This interaction uses an inactive Human MEK protein from Millipore, Cat no 14-420. braf-map2k1-9 26 32 unspecified role unspecified role bait bait P15056:p.Gly466Val mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain G V Wed May 02 15:08:13 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly466val' Tue Jun 30 16:13:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly466Val' P15056:p.Gly464Val mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain G V Wed May 02 17:09:32 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly464val' Tue Jun 30 19:00:14 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly464Val' region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P15056:p.Gly464Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain G E Wed May 02 17:38:56 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly464glu' Tue Jun 30 19:01:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly464Glu' 29 unspecified role unspecified role prey prey physical association physical association 4Bi map2k1-braf-7 14 30 unspecified role unspecified role bait bait 29 unspecified role unspecified role prey prey 32 unspecified role unspecified role prey prey association association Supp 1A This cell line only expressed mutated K-Ras (KRAS-MT). Coimmunoprecipitation (coIP) of MEK1 with BRAF and CRAF revealed that endogenous BRAF-MEK1 complexes are enriched in multiple KRAS-MT and BRAF-WT/KRAS-WT cell lines, but are significantly less abundant in the BRAF-V600E and BRAF-P loop cell lines. Only a weak band is seen for CRAF. map2k1-braf-8 12 29 unspecified role unspecified role prey prey 30 unspecified role unspecified role bait bait 32 unspecified role unspecified role prey prey association association Supp 1A This cell line only expressed mutated K-Ras (KRAS-MT). Coimmunoprecipitation (coIP) of MEK1 with BRAF and CRAF revealed that endogenous BRAF-MEK1 complexes are enriched in multiple KRAS-MT and BRAF-WT/KRAS-WT cell lines, but are significantly less abundant in the BRAF-V600E and BRAF-P loop cell lines. map2k1-braf-8 8 29 unspecified role unspecified role prey prey 30 unspecified role unspecified role bait bait 32 unspecified role unspecified role prey prey association association Supp 1A This cell line only expressed mutated K-Ras (KRAS-MT). Coimmunoprecipitation (coIP) of MEK1 with BRAF and CRAF revealed that endogenous BRAF-MEK1 complexes are enriched in multiple KRAS-MT and BRAF-WT/KRAS-WT cell lines, but are significantly less abundant in the BRAF-V600E and BRAF-P loop cell lines. Only a weak band is visible for CRAF. map2k1-braf-8 4 29 unspecified role unspecified role prey prey 30 unspecified role unspecified role bait bait 32 unspecified role unspecified role prey prey association association Supp 1A This cell line only expressed mutated K-Ras (KRAS-MT). Coimmunoprecipitation (coIP) of MEK1 with BRAF and CRAF revealed that endogenous BRAF-MEK1 complexes are enriched in multiple KRAS-MT and BRAF-WT/KRAS-WT cell lines, but are significantly less abundant in the BRAF-V600E and BRAF-P loop cell lines. Only a weak band is visible for CRAF. braf-map2k1-11 6 32 unspecified role unspecified role bait bait P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain V E Wed May 02 18:14:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 17:21:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' 30 unspecified role unspecified role prey prey 29 unspecified role unspecified role prey prey association association Supp 1A Coimmunoprecipitation (coIP) of MEK1 with BRAF and CRAF revealed that endogenous BRAF-MEK1 complexes are enriched in multiple KRAS-MT and BRAF-WT/KRAS-WT cell lines, but are significantly less abundant in the BRAF-V600E and BRAF-P loop cell lines. The MEK1 band is weak. map2k1-braf-9 13 30 unspecified role unspecified role bait bait 32 unspecified role unspecified role prey prey 29 unspecified role unspecified role prey prey association association 1A/C, 6C. Supp Fig 1. Coimmunoprecipitation (coIP) of MEK1 with BRAF and CRAF revealed that endogenous BRAF-MEK1 complexes are enriched in multiple KRAS-MT and BRAF-WT/KRAS-WT cell lines, but are significantly less abundant in the BRAF-V600E and BRAF-P loop cell lines. Vemurafenib - BRAF inhibitor effective only against cells with the V600E or V600 BRAF mutation. GDC-0879 -a selective B-Raf inhibitor raf1-braf-19 13 32 unspecified role unspecified role prey prey 30 unspecified role unspecified role prey prey 29 unspecified role unspecified role bait bait association association Supp Fig 1. Coimmunoprecipitation (coIP) of MEK1 with BRAF and CRAF revealed that endogenous BRAF-MEK1 complexes are enriched in multiple KRAS-MT and BRAF-WT/KRAS-WT cell lines, but are significantly less abundant in the BRAF-V600E and BRAF-P loop cell lines. The MEK1 band is weak. braf-map2k1-12 13 29 unspecified role unspecified role prey prey 32 unspecified role unspecified role bait bait 30 unspecified role unspecified role prey prey association association Supp Fig 1i Coimmunoprecipitation (coIP) of MEK1 with BRAF and CRAF revealed that endogenous BRAF-MEK1 complexes are enriched in multiple KRAS-MT and BRAF-WT/KRAS-WT cell lines, but are significantly less abundant in the BRAF-V600E and BRAF-P loop cell lines. map2k1-braf-10 24 30 unspecified role unspecified role bait bait 29 unspecified role unspecified role prey prey 32 unspecified role unspecified role prey prey association association Supp Fig 1C. When all MEK1 and MEK1-BRAF complexes are immunodepleted from cell lysates, free BRAF and CRAF remain in the flow through. braf-braf-30 23 32 unspecified role unspecified role prey prey region mcherry mcherry fluorescent protein tag mcherry undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P15056:p.Arg509His mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R H Wed May 02 14:52:54 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg509his' Tue Jun 30 18:48:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg509His' 32 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 30 unspecified role unspecified role prey prey association association Supp fig 1D. braf-mp2k1 22 30 unspecified role unspecified role neutral component neutral component kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) 32 unspecified role unspecified role neutral component neutral component kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. direct interaction direct interaction Supp Fig 2A braf-map2k1-13 22 32 unspecified role unspecified role neutral component neutral component kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P15056:p.Arg509His mutation mutation certain certain sequence position certain certain certain sequence position certain R H Wed May 02 18:01:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg509his' Tue Jun 30 19:32:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg509His' trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 30 unspecified role unspecified role neutral component neutral component kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) direct interaction direct interaction Supp Fig 2Ai braf-map2k1-14 22 30 unspecified role unspecified role neutral component neutral component 32 unspecified role unspecified role neutral component neutral component P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain V E Wed May 02 14:58:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 16:53:50 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. direct interaction direct interaction Supp Fig 2Aii braf-map2k1-16 20 30 unspecified role unspecified role prey prey c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 32 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P15056:p.Arg509His mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R H Wed May 02 19:57:40 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg509his' Tue Jun 30 20:25:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg509His' kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 228 224 direct interaction direct interaction Supp Fig 2B The authors have derived a Kd of ~40nM for BRAF-WT and ~60nM for the R509H BRAF mutant, but state that these differences were probably within experimental error. braf-map2k1-18 3 32 enzyme enzyme neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ser446 opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 30 enzyme target enzyme target substrate substrate neutral component neutral component ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Supp 2G, J,K, L + M Q02750 ser-218 opser purified recombinant BRAF (KD) is constitutively phosphorylated at S446. MEK was phosphorylated at other sites but these sites could not be unambiguously identified. (serine 605 or 607 and serine 298 or 299.) Q02750 ser-222 opser braf-map2k1-19 25 32 enzyme enzyme neutral component neutral component P15056:p.Arg462Glu mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain R E Wed May 02 17:16:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg462glu' Tue Jun 30 18:27:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg462Glu' P15056:p.Gly466Glu mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain G E Wed May 02 17:48:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly466glu' Tue Jun 30 17:38:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly466Glu' P15056:p.Gly466Val mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain G V Wed May 02 14:38:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly466val' Tue Jun 30 19:35:57 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly466Val' kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P15056:p.Gly464Glu mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain G E Wed May 02 16:18:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly464glu' Tue Jun 30 19:25:04 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly464Glu' P15056:p.Gly464Val mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain G V Wed May 02 18:04:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly464val' Tue Jun 30 19:52:51 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly464Val' trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 30 enzyme target enzyme target substrate substrate neutral component neutral component n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Supp 3B/C This interaction uses an inactive Human MEK protein from Millipore, Cat no 14-420. G464V and G464E have reduced enzymatic activity but similar Km(ATP) compared to BRAF-WT. atp-braf 5 31 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 32 unspecified role unspecified role neutral component neutral component tracer-178 alexa label alexa label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. direct interaction direct interaction Supp 4C The KD for ATP of BRAF-WT and the dimer defective BRAF R509H are similar. 2uM for WT and 6uM for mutant). braf-map2k1-20 18 30 unspecified role unspecified role prey prey n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) 32 unspecified role unspecified role bait bait P15056:p.[Arg462Glu;Ile666Arg] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R E certain certain sequence position certain certain certain sequence position certain I R Wed May 02 15:26:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg462glu,ile666arg' Tue Jun 30 19:44:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg462Glu;Ile666Arg]' region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P15056:p.Gly464Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain G E Wed May 02 19:21:47 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly464glu' Tue Jun 30 16:47:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly464Glu' P15056:p.Gly466Val mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain G V Wed May 02 17:34:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly466val' Tue Jun 30 16:18:34 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly466Val' P15056:p.Gly464Val mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain G V Wed May 02 15:22:41 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly464val' Tue Jun 30 15:52:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly464Val' P15056:p.Arg462Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R E Wed May 02 15:00:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg462glu' Tue Jun 30 19:32:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg462Glu' physical association physical association 4B top,4D map2k1-braf-11 10 32 unspecified role unspecified role prey prey kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 30 unspecified role unspecified role bait bait n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range phosphores phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) direct interaction direct interaction Supp fig 5B phosres added as a disrupting ptm - at particpant level. Vemurafenib - a BRAF inhibitor GDC-0879 - a BRAF inhibitor