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/ http://www.ebi.ac.uk/ haling-2014-1 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-22 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-5 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-7 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-8 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-9 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-11 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-12 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-14 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-2 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-3 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-4 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-6 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-10 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-13 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-15 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-16 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-17 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-18 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-20 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-21 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-23 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-24 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-25 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-26 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-27 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation haling-2014-28 Structure of the BRAF-MEK Complex Reveals a Kinase Activity Independent Role for BRAF in MAPK Signaling. 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.. 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 Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation 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 raf1_human RAF proto-oncogene serine/threonine-protein kinase RAF1 RAF Proto-oncogene c-RAF Raf-1 protein protein human Homo sapiens Human MEHIQGAWKTISNGFGFKDAVFDGSSCISPTIVQQFGYQRRASDDGKLTDPSKTSNTIRVFLPNKQRTVVNVRNGMSLHDCLMKALKVRGLQPECCAVFRLLHEHKGKKARLDWNTDAASLIGEELQVDFLDHVPLTTHNFARKTFLKLAFCDICQKFLLNGFRCQTCGYKFHEHCSTKVPTMCVDWSNIRQLLLFPNSTIGDSGVPALPSLTMRRMRESVSRMPVSSQHRYSTPHAFTFNTSSPSSEGSLSQRQRSTSTPNVHMVSTTLPVDSRMIEDAIRSHSESASPSALSSSPNNLSPTGWSQPKTPVPAQRERAPVSGTQEKNKIRPRGQRDSSYYWEIEASEVMLSTRIGSGSFGTVYKGKWHGDVAVKILKVVDPTPEQFQAFRNEVAVLRKTRHVNILLFMGYMTKDNLAIVTQWCEGSSLYKHLHVQETKFQMFQLIDIARQTAQGMDYLHAKNIIHRDMKSNNIFLHEGLTVKIGDFGLATVKSRWSGSQQVEQPTGSVLWMAPEVIRMQDNNPFSFQSDVYSYGIVLYELMTGELPYSHINNRDQIIFMVGRGYASPDLSKLYKNCPKAMKRLVADCVKKVKEERPLFPQILSSIELLQHSLPKINRSASEPSLHRAAHTEDINACTLTTSPRLPVF 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 g-573 G-573 small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) 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 gdc-0879 GDC-0879 small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) 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) map2k1-braf 1574092 n/a 1574119 unspecified role unspecified role prey prey n/a 1574120 unspecified role unspecified role prey prey n/a 1574121 unspecified role unspecified role bait bait association association false false false 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-9 1574092 n/a 1574121 unspecified role unspecified role bait bait n/a 1574119 unspecified role unspecified role prey prey n/a 1574120 unspecified role unspecified role prey prey association association false false false 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 1574092 n/a 1574119 unspecified role unspecified role prey prey n/a 1574121 unspecified role unspecified role prey prey n/a 1574120 unspecified role unspecified role bait bait association association false false false 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 1574092 n/a 1574120 unspecified role unspecified role prey prey n/a 1574119 unspecified role unspecified role bait bait n/a 1574121 unspecified role unspecified role prey prey association association false false false 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. braf-mp2k1 1574093 n/a 1574121 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 false 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 false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 n/a 1574119 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 false 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 false 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. Stoichiometry: 2.0 direct interaction direct interaction false false false Supp Fig 2A braf-map2k1-13 1574093 n/a 1574119 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 false 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 false P15056:p.Arg509His mutation mutation certain certain sequence position certain certain certain sequence position certain false 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. Stoichiometry: 1.0 n/a 1574121 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 false 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 false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 direct interaction direct interaction false false false Supp Fig 2Ai braf-map2k1-14 1574093 n/a 1574121 unspecified role unspecified role neutral component neutral component Stoichiometry: 2.0 n/a 1574119 unspecified role unspecified role neutral component neutral component P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false 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 false kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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. Stoichiometry: 2.0 direct interaction direct interaction false false false Supp Fig 2Aii map2k1-braf-2 1574094 n/a 1574119 unspecified role unspecified role prey prey n/a 1574120 unspecified role unspecified role prey prey n/a 1574121 unspecified role unspecified role bait bait association association false false false 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. braf-map2k1-3 1574094 n/a 1574121 unspecified role unspecified role prey prey n/a 1574119 unspecified role unspecified role bait bait association association false false false 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. map2k1-braf-4 1574095 n/a 1574120 unspecified role unspecified role prey prey n/a 1574121 unspecified role unspecified role bait bait n/a 1574119 unspecified role unspecified role prey prey P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false 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. braf-map2k1-11 1574095 n/a 1574119 unspecified role unspecified role bait bait P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false n/a 1574121 unspecified role unspecified role prey prey n/a 1574120 unspecified role unspecified role prey prey association association false false false 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-5 1574096 n/a 1574120 unspecified role unspecified role prey prey n/a 1574119 unspecified role unspecified role prey prey P15056:p.Gly464Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false n/a 1574121 unspecified role unspecified role bait bait association association false false false 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. raf1-braf-15 1574096 n/a 1574120 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 false n/a 1574121 unspecified role unspecified role prey prey n/a 1574119 unspecified role unspecified role prey prey P15056:p.Gly464Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false 1B map2k1-braf-6 1574097 n/a 1574119 unspecified role unspecified role prey prey P15056:p.Gly464Val mutation mutation certain certain sequence position certain certain certain sequence position certain false n/a 1574121 unspecified role unspecified role bait bait n/a 1574120 unspecified role unspecified role prey prey association association false false false 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-16 1574097 n/a 1574119 unspecified role unspecified role prey prey P15056:p.Gly464Val mutation mutation certain certain sequence position certain certain certain sequence position certain false n/a 1574120 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 false n/a 1574121 unspecified role unspecified role prey prey association association false false false 1B adenosine_5___beta_gamma_methylene_triphosphate-braf-1 1574098 n/a 1574122 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1574123 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1574121 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 false kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 n/a 1574119 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 false kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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. Stoichiometry: 2.0 1574210 1574207 direct interaction direct interaction false false false Table S1, (figure 2) 2.8A working 18.64%, Free: 24.02% braf-gdc_0879 1574098 n/a 1574124 unspecified role unspecified role neutral component neutral component n/a 1574119 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 false P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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. Stoichiometry: 2.0 direct interaction direct interaction false false false Table S1 (fig 2) 2.8A working:21.46%, Free: 27.45%. braf-map2k1-4 1574099 n/a 1574119 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 false P15056:p.Arg462Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P15056:p.[Arg462Glu;Ile617Arg] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P15056:p.Ile617Arg mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false n/a 1574121 unspecified role unspecified role prey prey physical association physical association false false false 3B braf-map2k1-9 1574099 n/a 1574119 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 false P15056:p.Gly464Val mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false 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 false P15056:p.Gly464Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false n/a 1574120 unspecified role unspecified role prey prey physical association physical association false false false 4Bi braf-map2k1-6 1574100 n/a 1574121 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 false kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q02750:p.Ile310Glu mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false Q02750:p.Ser228Glu mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false Q02750:p.[Ser228Glu;Met230Ala] mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q02750:p.[Ile310Glu;Leu314Glu] mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q02750:p.Leu314Glu mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1574119 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 false kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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. 1574243 1574234 phosphorylation phosphorylation reaction phosphorylation false false false 3Di P15056 increases activity of Q02750 braf-map2k1-7 1574100 n/a 1574121 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 false 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 false ecoli Escherichia coli (strain K12) n/a 1574119 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 false kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false false false 3Dii This interaction uses an inactive Human MEK protein from Millipore, Cat no 14-420. map2k1-braf-1 1574101 n/a 1574119 unspecified role unspecified role prey prey n/a 1574121 unspecified role unspecified role bait bait n/a 1574120 unspecified role unspecified role prey prey association association false false false 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 1574102 n/a 1574120 unspecified role unspecified role prey prey n/a 1574121 unspecified role unspecified role bait bait n/a 1574119 unspecified role unspecified role prey prey association association false false false 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 1574103 n/a 1574121 unspecified role unspecified role bait bait n/a 1574120 unspecified role unspecified role prey prey n/a 1574119 unspecified role unspecified role prey prey association association false false false 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-3 1574104 n/a 1574119 unspecified role unspecified role prey prey n/a 1574120 unspecified role unspecified role prey prey n/a 1574121 unspecified role unspecified role bait bait association association false false false 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. raf1-braf-17 1574105 n/a 1574121 unspecified role unspecified role prey prey n/a 1574119 unspecified role unspecified role prey prey n/a 1574120 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 false association association false false false 1B, 6E. GDC-0879 -a selective B-Raf inhibitor braf-map2k1-5 1574106 n/a 1574121 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 false 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 false Q02750:p.[Ser228Glu;Met230Ala] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q02750:p.Ile310Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q02750:p.Leu314Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q02750:p.Ser228Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q02750:p.[Ile310Glu;Leu314Glu] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1574119 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 false 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 false 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. 1574283 1574274 direct interaction direct interaction false false false 3C, 6B, Supp 2E G-573 -a MEK inhibitor, GDC-0879 -a BRAF inhibitor, PLX-4720-a BRAF inhibitor. ATP map2k1-braf-7 1574107 n/a 1574121 unspecified role unspecified role bait bait n/a 1574120 unspecified role unspecified role prey prey n/a 1574119 unspecified role unspecified role prey prey association association false false false 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 1574108 n/a 1574120 unspecified role unspecified role prey prey n/a 1574121 unspecified role unspecified role bait bait n/a 1574119 unspecified role unspecified role prey prey association association false false false 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 1574109 n/a 1574120 unspecified role unspecified role prey prey n/a 1574121 unspecified role unspecified role bait bait n/a 1574119 unspecified role unspecified role prey prey association association false false false 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 1574110 n/a 1574120 unspecified role unspecified role prey prey n/a 1574121 unspecified role unspecified role bait bait n/a 1574119 unspecified role unspecified role prey prey association association false false false 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-10 1574111 n/a 1574121 unspecified role unspecified role bait bait n/a 1574120 unspecified role unspecified role prey prey n/a 1574119 unspecified role unspecified role prey prey association association false false false 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 1574112 n/a 1574119 unspecified role unspecified role prey prey region mcherry mcherry fluorescent protein tag mcherry undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P15056:p.Arg509His mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false n/a 1574119 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 false n/a 1574121 unspecified role unspecified role prey prey association association false false false Supp fig 1D. braf-map2k1-16 1574113 n/a 1574121 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 false kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 n/a 1574119 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 false P15056:p.Arg509His mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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. Stoichiometry: 1.0 1574319 1574315 direct interaction direct interaction false false false 1574113 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 1574114 n/a 1574119 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 false kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false 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. n/a 1574121 enzyme target enzyme target substrate substrate neutral component neutral component ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false 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 1574115 n/a 1574119 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 false P15056:p.Gly466Glu mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain false P15056:p.Gly466Val mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain false kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false P15056:p.Gly464Glu mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false P15056:p.Gly464Val mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false 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. n/a 1574121 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 false 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 false ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false 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 1574116 n/a 1574125 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1574119 unspecified role unspecified role neutral component neutral component tracer-178 alexa label alexa label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false 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 false false false 1574116 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 1574117 n/a 1574121 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 false 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 false ecoli Escherichia coli (strain K12) n/a 1574119 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 false certain certain sequence position certain certain certain sequence position certain false 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 false P15056:p.Gly464Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P15056:p.Gly466Val mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P15056:p.Gly464Val mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P15056:p.Arg462Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false 4B top,4D map2k1-braf-11 1574118 n/a 1574119 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 false 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 false 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. n/a 1574121 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 false 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 false 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 false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Supp fig 5B Vemurafenib - a BRAF inhibitor GDC-0879 - a BRAF inhibitor