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/ moriceau-2015-4 human-m249parent Human M249-Melanoma cell line with Braf V600E mutation Human m249parent M249-Melanoma cell line with Braf V600E mutation anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Moriceau G., Hugo W., Hong A., Shi H., Kong X., Yu CC., Koya RC., Samatar AA., Khanlou N., Braun J., Ruchalski K., Seifert H., Larkin J., Dahlman KB., Johnson DB., Algazi A., Sosman JA., Ribas A., Lo RS. Cancer - Interactions investigated in the context of cancer Figure 5A: IP results using the parent cell line M249 have not been included as a non-specific antibody was used to detect both MEK 1 and 2; these proteins cannot be distinguished as they have a near identical M.Wt. The authors have confirmed that all constructs are human in origin. rlo@mednet.ucla.edu Cancer cell 2015 Only protein-protein interactions imex curation imex curation moriceau-2015-5 human-m249parent Human M249-Melanoma cell line with Braf V600E mutation Human m249parent M249-Melanoma cell line with Braf V600E mutation anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Moriceau G., Hugo W., Hong A., Shi H., Kong X., Yu CC., Koya RC., Samatar AA., Khanlou N., Braun J., Ruchalski K., Seifert H., Larkin J., Dahlman KB., Johnson DB., Algazi A., Sosman JA., Ribas A., Lo RS. Cancer - Interactions investigated in the context of cancer Anti-BRAF signals detected in the FLAG immunoprecipitates presumably contained both endogenous and exogenous V600E BRAF. The authors have confirmed that all constructs are human in origin. rlo@mednet.ucla.edu Cancer cell 2015 Only protein-protein interactions imex curation imex curation moriceau-2015-1 human-ddr4 Human DDR4- single cell derived sub-clone of melanoma M249 cell line;"drug-addicted" - resistant to Braf and MEK inhibitors, and exhibits loss of viability in their absence. Human ddr4 DDR4- single cell sub-clone of M249 cell line;"drug-addicted" - resistant to Braf and MEK inhibitors, and loss of viability in absence of drug. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Moriceau G., Hugo W., Hong A., Shi H., Kong X., Yu CC., Koya RC., Samatar AA., Khanlou N., Braun J., Ruchalski K., Seifert H., Larkin J., Dahlman KB., Johnson DB., Algazi A., Sosman JA., Ribas A., Lo RS. Cancer - Interactions investigated in the context of cancer Figure 5A: IP results using the parent cell line M249 have not been included as a non-specific antibody was used to detect both MEK 1 and 2; these proteins cannot be distinguished as they have a near identical M.Wt. The authors have confirmed that all constructs are human in origin. rlo@mednet.ucla.edu Cancer cell 2015 Only protein-protein interactions imex curation imex curation moriceau-2015-2 human-m249parent Human M249-Melanoma cell line with Braf V600E mutation Human m249parent M249-Melanoma cell line with Braf V600E mutation anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Moriceau G., Hugo W., Hong A., Shi H., Kong X., Yu CC., Koya RC., Samatar AA., Khanlou N., Braun J., Ruchalski K., Seifert H., Larkin J., Dahlman KB., Johnson DB., Algazi A., Sosman JA., Ribas A., Lo RS. Cancer - Interactions investigated in the context of cancer Figure 5A: IP results using the parent cell line M249 have not been included as a non-specific antibody was used to detect both MEK 1 and 2; these proteins cannot be distinguished as they have a near identical M.Wt. The authors have confirmed that all constructs are human in origin. rlo@mednet.ucla.edu Cancer cell 2015 Only protein-protein interactions imex curation imex curation moriceau-2015-3 human-ddr5 Human DDR5- single cell derived sub-clone of melanoma M249 cell line;"drug-addicted" - resistant to Braf and MEK inhibitors, and exhibits loss of viability in their absence. Human ddr5 Human DDR5- single cell sub-clone of melanoma M249 cell line;"drug-addicted" - resistant to Braf and MEK inhibitors, and exhibits loss of viability in absence of these inhibitors. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Moriceau G., Hugo W., Hong A., Shi H., Kong X., Yu CC., Koya RC., Samatar AA., Khanlou N., Braun J., Ruchalski K., Seifert H., Larkin J., Dahlman KB., Johnson DB., Algazi A., Sosman JA., Ribas A., Lo RS. Cancer - Interactions investigated in the context of cancer Figure 5A: IP results using the parent cell line M249 have not been included as a non-specific antibody was used to detect both MEK 1 and 2; these proteins cannot be distinguished as they have a near identical M.Wt. The authors have confirmed that all constructs are human in origin. rlo@mednet.ucla.edu Cancer cell 2015 Only protein-protein interactions imex curation 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 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 mp2k2_human Dual specificity mitogen-activated protein kinase kinase 2 MAP2K2 MEK2 MKK2 PRKMK2 ERK activator kinase 2 MAPK/ERK kinase 2 protein protein human Homo sapiens Human MLARRKPVLPALTINPTIAEGPSPTSEGASEANLVDLQKKLEELELDEQQKKRLEAFLTQKAKVGELKDDDFERISELGAGNGGVVTKVQHRPSGLIMARKLIHLEIKPAIRNQIIRELQVLHECNSPYIVGFYGAFYSDGEISICMEHMDGGSLDQVLKEAKRIPEEILGKVSIAVLRGLAYLREKHQIMHRDVKPSNILVNSRGEIKLCDFGVSGQLIDSMANSFVGTRSYMAPERLQGTHYSVQSDIWSMGLSLVELAVGRYPIPPPDAKELEAIFGRPVVDGEEGEPHSISPRPRPPGRPVSGHGMDSRPAMAIFELLDYIVNEPPPKLPNGVFTPDFQEFVNKCLIKNPAERADLKMLTNHTFIKRSEVEEVDFAGWLCKTLRLNQPGTPTRTAV map2k1-braf-18 1800404 N/A 1800409 unspecified role unspecified role prey prey N/A ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope 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 P15056:p.Arg662Leu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P15056:p.Val600Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false N/A 1800410 unspecified role unspecified role bait bait Q02750:p.Phe129Leu mutation mutation certain certain sequence position certain certain certain sequence position certain false N/A 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 physical association physical association false false false 5F,6D Both the BRAF (WT) and BRAF V600E mutant reacted preferentially to the MEK1 F129L and Q56P mutants, rather than to WT. map2k1-braf-19 1800404 N/A 1800409 unspecified role unspecified role prey prey N/A ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P15056:p.Val600Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain 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 P15056:p.Arg662Leu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false N/A 1800410 unspecified role unspecified role bait bait Q02750:p.Gln56Pro mutation mutation certain certain sequence position certain certain certain sequence position certain false N/A 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 physical association physical association false false false 5F, 6D Both the BRAF (WT) and BRAF V600E mutant reacted preferentially to the MEK1 F129L and Q56P mutants, rather than to WT. map2k1-braf-20 1800404 N/A 1800409 unspecified role unspecified role prey prey N/A ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P15056:p.Val600Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false N/A 1800410 unspecified role unspecified role bait bait Q02750:p.Glu203Lys mutation mutation certain certain sequence position certain certain certain sequence position certain false N/A 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 physical association physical association false false false Supp 4 map2k1-braf-21 1800404 N/A 1800409 unspecified role unspecified role prey prey N/A ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope 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 N/A 1800410 unspecified role unspecified role bait bait Q02750:p.Lys59del mutation increasing mutation increasing interaction mutation increasing deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false N/A 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 physical association physical association false false false Supp 4i An in-frame deletion mutant (K59) showed binding to the BRAF V600E mutant but not the WT. map2k1-braf-22 1800405 N/A 1800409 unspecified role unspecified role prey prey N/A ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope 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 N/A 1800410 unspecified role unspecified role bait bait Q02750:p.Lys59del mutation increasing mutation increasing interaction mutation increasing deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false N/A 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 physical association physical association false false false Supp 4i An in-frame deletion mutant (K59) showed binding to the BRAF V600E mutant but not the WT. map2k1-braf-23 1800405 N/A 1800409 unspecified role unspecified role prey prey N/A ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P15056:p.Val600Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain 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 P15056:p.Arg662Leu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false N/A 1800410 unspecified role unspecified role bait bait Q02750:p.Gln56Pro mutation mutation certain certain sequence position certain certain certain sequence position certain false N/A 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 physical association physical association false false false 5F, 6D Both the BRAF (WT) and BRAF V600E mutant reacted preferentially to the MEK1 F129L and Q56P mutants, rather than to WT. map2k1-braf-24 1800405 N/A 1800409 unspecified role unspecified role prey prey N/A ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope 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 P15056:p.Arg662Leu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P15056:p.Val600Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false N/A 1800410 unspecified role unspecified role bait bait Q02750:p.Phe129Leu mutation mutation certain certain sequence position certain certain certain sequence position certain false N/A 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 physical association physical association false false false 5F,6D Both the BRAF (WT) and BRAF V600E mutant reacted preferentially to the MEK1 F129L and Q56P mutants, rather than to WT. map2k1-braf-25 1800405 N/A 1800409 unspecified role unspecified role prey prey N/A ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope 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 N/A 1800410 unspecified role unspecified role bait bait Q02750:p.Glu203Lys mutation mutation certain certain sequence position certain certain certain sequence position certain false N/A 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 physical association physical association false false false Supp 4 map2k1-braf-12 1800406 N/A 1800409 unspecified role unspecified role prey prey P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false N/A 1800410 unspecified role unspecified role bait bait physical association physical association false false false 5B map2k1-braf-16 1800406 N/A 1800409 unspecified role unspecified role prey prey P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false N/A 1800411 unspecified role unspecified role bait bait physical association physical association false false false 5C map2k1-braf-13 1800407 N/A 1800409 unspecified role unspecified role prey prey P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false N/A 1800410 unspecified role unspecified role bait bait physical association physical association false false false 5B map2k1-braf-17 1800407 N/A 1800409 unspecified role unspecified role prey prey P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false N/A 1800411 unspecified role unspecified role bait bait physical association physical association false false false 5C map2k1-braf-14 1800408 N/A 1800409 unspecified role unspecified role prey prey P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false N/A 1800410 unspecified role unspecified role bait bait Q02750:p.Phe129Leu mutation mutation certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false 5B More BRAF was detected bound to MEK1 in the DDR-5 cell line than in DDR-4 and parental M249 lines. map2k1-braf-15 1800408 N/A 1800411 unspecified role unspecified role bait bait N/A 1800409 unspecified role unspecified role prey prey P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false 5C