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/ Tunable-combinatorial mechanisms of acquired resistance limit the efficacy of BRAF/MEK cotargeting but result in melanoma drug addiction. Tunable-combinatorial mechanisms of acquired resistance limit the efficacy of BRAF/MEK cotargeting but result in melanoma drug addiction. Cancer cell 2015 imex curation 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 Fig5C - I have not added any PTM to Bait (MEK2 phosphorylation) as it is not clear if phospho or non-phospho version, or both, is binding and we cannot add as "possible" ptm. (short label not visible) The authors have confirmed that all constructs are human in origin. rlo@mednet.ucla.edu Only protein-protein interactions imex curation 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. Fig5C - I have not added any PTM to Bait (MEK2 phosphorylation) as it is not clear if phospho or non-phospho version, or both, is binding and we cannot add as "possible" ptm. (short label not visible) The authors have confirmed that all constructs are human in origin. rlo@mednet.ucla.edu Accepted 2015-JUN-17 by ORCHARD Cancer cell 2015 Only protein-protein interactions imex curation imex curation EBI-10822129 - Fig. 6F:- there is no WT for (IP= FLAG) FLAG-MEK1 reacting -as it does not react - the WT is for BRAF only. (EBI-10822108) this interaction shows the F129L mutant as Bait. EBI-10823053:- also Bait is FLAG-MEK1 - which is the E203K mutant (no WT) and the V600E is the Prey - BRAF, which does show the WT. EBI-10823216:- I had another idea - according to "Standard Ambiguity Codes" (IUPAC)- a deletion is represented by a "." - a "dot", so I have entered this as the 'Resulting sequence', (despite never having come across this before). I suppose the alternative is to do as you suggest- enter range of 59-393 for original sequence and then the sequence for amino acids 60-393 for the Resultant sequence? ; EBI-10823286- I have altered the BRAF "mutation increasing" to just "mutation". I have entered a "." for the MEK1 mutation, as in the previous Expt, as this is less clunky than the alternative - if you dont agree/like this please send this publication back. Tunable-combinatorial mechanisms of acquired resistance limit the efficacy of BRAF/MEK cotargeting but result in melanoma drug addiction. Tunable-combinatorial mechanisms of acquired resistance limit the efficacy of BRAF/MEK cotargeting but result in melanoma drug addiction. Cancer cell 2015 imex curation 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 Fig5C - I have not added any PTM to Bait (MEK2 phosphorylation) as it is not clear if phospho or non-phospho version, or both, is binding and we cannot add as "possible" ptm. (short label not visible) The authors have confirmed that all constructs are human in origin. rlo@mednet.ucla.edu Only protein-protein interactions imex curation 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. Fig5C - I have not added any PTM to Bait (MEK2 phosphorylation) as it is not clear if phospho or non-phospho version, or both, is binding and we cannot add as "possible" ptm. (short label not visible) The authors have confirmed that all constructs are human in origin. rlo@mednet.ucla.edu Accepted 2015-JUN-17 by ORCHARD Cancer cell 2015 Only protein-protein interactions imex curation imex curation EBI-10822129 - Fig. 6F:- there is no WT for (IP= FLAG) FLAG-MEK1 reacting -as it does not react - the WT is for BRAF only. (EBI-10822108) this interaction shows the F129L mutant as Bait. EBI-10823053:- also Bait is FLAG-MEK1 - which is the E203K mutant (no WT) and the V600E is the Prey - BRAF, which does show the WT. EBI-10823216:- I had another idea - according to "Standard Ambiguity Codes" (IUPAC)- a deletion is represented by a "." - a "dot", so I have entered this as the 'Resulting sequence', (despite never having come across this before). I suppose the alternative is to do as you suggest- enter range of 59-393 for original sequence and then the sequence for amino acids 60-393 for the Resultant sequence? ; EBI-10823286- I have altered the BRAF "mutation increasing" to just "mutation". I have entered a "." for the MEK1 mutation, as in the previous Expt, as this is less clunky than the alternative - if you dont agree/like this please send this publication back. Tunable-combinatorial mechanisms of acquired resistance limit the efficacy of BRAF/MEK cotargeting but result in melanoma drug addiction. Tunable-combinatorial mechanisms of acquired resistance limit the efficacy of BRAF/MEK cotargeting but result in melanoma drug addiction. Cancer cell 2015 imex curation 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 Fig5C - I have not added any PTM to Bait (MEK2 phosphorylation) as it is not clear if phospho or non-phospho version, or both, is binding and we cannot add as "possible" ptm. (short label not visible) The authors have confirmed that all constructs are human in origin. rlo@mednet.ucla.edu Only protein-protein interactions imex curation 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. Fig5C - I have not added any PTM to Bait (MEK2 phosphorylation) as it is not clear if phospho or non-phospho version, or both, is binding and we cannot add as "possible" ptm. (short label not visible) The authors have confirmed that all constructs are human in origin. rlo@mednet.ucla.edu Accepted 2015-JUN-17 by ORCHARD Cancer cell 2015 Only protein-protein interactions imex curation imex curation EBI-10822129 - Fig. 6F:- there is no WT for (IP= FLAG) FLAG-MEK1 reacting -as it does not react - the WT is for BRAF only. (EBI-10822108) this interaction shows the F129L mutant as Bait. EBI-10823053:- also Bait is FLAG-MEK1 - which is the E203K mutant (no WT) and the V600E is the Prey - BRAF, which does show the WT. EBI-10823216:- I had another idea - according to "Standard Ambiguity Codes" (IUPAC)- a deletion is represented by a "." - a "dot", so I have entered this as the 'Resulting sequence', (despite never having come across this before). I suppose the alternative is to do as you suggest- enter range of 59-393 for original sequence and then the sequence for amino acids 60-393 for the Resultant sequence? ; EBI-10823286- I have altered the BRAF "mutation increasing" to just "mutation". I have entered a "." for the MEK1 mutation, as in the previous Expt, as this is less clunky than the alternative - if you dont agree/like this please send this publication back. Tunable-combinatorial mechanisms of acquired resistance limit the efficacy of BRAF/MEK cotargeting but result in melanoma drug addiction. Tunable-combinatorial mechanisms of acquired resistance limit the efficacy of BRAF/MEK cotargeting but result in melanoma drug addiction. Cancer cell 2015 imex curation 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 Fig5C - I have not added any PTM to Bait (MEK2 phosphorylation) as it is not clear if phospho or non-phospho version, or both, is binding and we cannot add as "possible" ptm. (short label not visible) The authors have confirmed that all constructs are human in origin. rlo@mednet.ucla.edu Only protein-protein interactions imex curation 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. Fig5C - I have not added any PTM to Bait (MEK2 phosphorylation) as it is not clear if phospho or non-phospho version, or both, is binding and we cannot add as "possible" ptm. (short label not visible) The authors have confirmed that all constructs are human in origin. rlo@mednet.ucla.edu Accepted 2015-JUN-17 by ORCHARD Cancer cell 2015 Only protein-protein interactions imex curation imex curation EBI-10822129 - Fig. 6F:- there is no WT for (IP= FLAG) FLAG-MEK1 reacting -as it does not react - the WT is for BRAF only. (EBI-10822108) this interaction shows the F129L mutant as Bait. EBI-10823053:- also Bait is FLAG-MEK1 - which is the E203K mutant (no WT) and the V600E is the Prey - BRAF, which does show the WT. EBI-10823216:- I had another idea - according to "Standard Ambiguity Codes" (IUPAC)- a deletion is represented by a "." - a "dot", so I have entered this as the 'Resulting sequence', (despite never having come across this before). I suppose the alternative is to do as you suggest- enter range of 59-393 for original sequence and then the sequence for amino acids 60-393 for the Resultant sequence? ; EBI-10823286- I have altered the BRAF "mutation increasing" to just "mutation". I have entered a "." for the MEK1 mutation, as in the previous Expt, as this is less clunky than the alternative - if you dont agree/like this please send this publication back. Tunable-combinatorial mechanisms of acquired resistance limit the efficacy of BRAF/MEK cotargeting but result in melanoma drug addiction. Tunable-combinatorial mechanisms of acquired resistance limit the efficacy of BRAF/MEK cotargeting but result in melanoma drug addiction. Cancer cell 2015 imex curation 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 Fig5C - I have not added any PTM to Bait (MEK2 phosphorylation) as it is not clear if phospho or non-phospho version, or both, is binding and we cannot add as "possible" ptm. (short label not visible) The authors have confirmed that all constructs are human in origin. rlo@mednet.ucla.edu Only protein-protein interactions imex curation 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. Fig5C - I have not added any PTM to Bait (MEK2 phosphorylation) as it is not clear if phospho or non-phospho version, or both, is binding and we cannot add as "possible" ptm. (short label not visible) The authors have confirmed that all constructs are human in origin. rlo@mednet.ucla.edu Accepted 2015-JUN-17 by ORCHARD Cancer cell 2015 Only protein-protein interactions imex curation imex curation mp2k1_human Dual specificity mitogen-activated protein kinase kinase 1 ERK activator kinase 1 MAPK/ERK kinase 1 MAP2K1 MEK1 PRKMK1 protein protein human Homo sapiens Human MPKKKPTPIQLNPAPDGSAVNGTSSAETNLEALQKKLEELELDEQQRKRLEAFLTQKQKVGELKDDDFEKISELGAGNGGVVFKVSHKPSGLVMARKLIHLEIKPAIRNQIIRELQVLHECNSPYIVGFYGAFYSDGEISICMEHMDGGSLDQVLKKAGRIPEQILGKVSIAVIKGLTYLREKHKIMHRDVKPSNILVNSRGEIKLCDFGVSGQLIDSMANSFVGTRSYMSPERLQGTHYSVQSDIWSMGLSLVEMAVGRYPIPPPDAKELELMFGCQVEGDAAETPPRPRTPGRPLSSYGMDSRPPMAIFELLDYIVNEPPPKLPSGVFSLEFQDFVNKCLIKNPAERADLKQLMVHAFIKRSDAEEVDFAGWLCSTIGLNQPSTPTHAAGV DIP protein Q02750 original sequence version: 96 DIP protein Q02750 original sequence version: 93 DIP protein Q02750-1 original sequence version: 151 DIP protein Q02750 original sequence version: 151 DIP protein Q02750-1 original sequence version: 166 0344118FFC842D51 braf_human Serine/threonine-protein kinase B-raf BRAF BRAF1 RAFB1 v-Raf murine sarcoma viral oncogene homolog B1 p94 Proto-oncogene B-Raf protein protein human Homo sapiens Human MAALSGGGGGGAEPGQALFNGDMEPEAGAGAGAAASSAADPAIPEEVWNIKQMIKLTQEHIEALLDKFGGEHNPPSIYLEAYEEYTSKLDALQQREQQLLESLGNGTDFSVSSSASMDTVTSSSSSSLSVLPSSLSVFQNPTDVARSNPKSPQKPIVRVFLPNKQRTVVPARCGVTVRDSLKKALMMRGLIPECCAVYRIQDGEKKPIGWDTDISWLTGEELHVEVLENVPLTTHNFVRKTFFTLAFCDFCRKLLFQGFRCQTCGYKFHQRCSTEVPLMCVNYDQLDLLFVSKFFEHHPIPQEEASLAETALTSGSSPSAPASDSIGPQILTSPSPSKSIPIPQPFRPADEDHRNQFGQRDRSSSAPNVHINTIEPVNIDDLIRDQGFRGDGGSTTGLSATPPASLPGSLTNVKALQKSPGPQRERKSSSSSEDRNRMKTLGRRDSSDDWEIPDGQITVGQRIGSGSFGTVYKGKWHGDVAVKMLNVTAPTPQQLQAFKNEVGVLRKTRHVNILLFMGYSTKPQLAIVTQWCEGSSLYHHLHIIETKFEMIKLIDIARQTAQGMDYLHAKSIIHRDLKSNNIFLHEDLTVKIGDFGLATVKSRWSGSHQFEQLSGSILWMAPEVIRMQDKNPYSFQSDVYAFGIVLYELMTGQLPYSNINNRDQIIFMVGRGYLSPDLSKVRSNCPKAMKRLMAECLKKKRDERPLFPQILASIELLARSLPKIHRSASEPSLNRAGFQTEDFSLYACASPKTPIQAGGYGAFPVH DIP protein P15056 original sequence version: 182 DIP protein P15056 original sequence version: 133 DIP protein P15056 original sequence version: 147 DIP protein P15056 original sequence version: 173 DIP protein P15056 original sequence version: 178 DIP protein P15056 original sequence version: 188 DIP protein P15056 original sequence version: 193 DIP protein P15056 original sequence version: 195 DIP protein P15056 original sequence version: 146 DIP protein P15056 original sequence version: 153 0798C2AAB487E813 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 3401D522515C30A5 map2k1-braf-12 1 7 unspecified role unspecified role prey prey P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain V E Wed May 02 19:45:20 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 17:18:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' 6 unspecified role unspecified role bait bait physical association physical association 5B map2k1-braf-13 2 7 unspecified role unspecified role prey prey P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain V E Wed May 02 16:05:57 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 20:06:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' 6 unspecified role unspecified role bait bait physical association physical association 5B map2k1-braf-14 3 7 unspecified role unspecified role prey prey P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain V E Wed May 02 15:55:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 16:25:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' 6 unspecified role unspecified role bait bait Q02750:p.Phe129Leu mutation mutation certain certain sequence position certain certain certain sequence position certain F L Wed May 02 19:49:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe129leu' Tue Jun 30 18:55:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe129Leu' physical association physical association 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 3 8 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain V E Wed May 02 14:28:48 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 18:54:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' physical association physical association 5C map2k1-braf-16 1 7 unspecified role unspecified role prey prey P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain V E Wed May 02 20:12:06 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 19:24:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' 8 unspecified role unspecified role bait bait physical association physical association 5C map2k1-braf-17 2 7 unspecified role unspecified role prey prey P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain V E Wed May 02 16:32:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 19:15:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' 8 unspecified role unspecified role bait bait physical association physical association 5C map2k1-braf-18 4 7 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 P15056:p.Arg509His mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R H Wed May 02 19:17:04 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg509his' Tue Jun 30 16:26:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg509His' P15056:p.Arg662Leu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R L Wed May 02 18:37:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg662leu' Tue Jun 30 17:14:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg662Leu' P15056:p.Val600Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain V E Wed May 02 15:17:42 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 19:28:44 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' 6 unspecified role unspecified role bait bait Q02750:p.Phe129Leu mutation mutation certain certain sequence position certain certain certain sequence position certain F L Wed May 02 16:57:56 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe129leu' Tue Jun 30 18:11:58 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe129Leu' 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 physical association physical association 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 4 7 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 P15056:p.Val600Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain V E Wed May 02 18:36:59 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 17:37:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' P15056:p.Arg509His mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R H Wed May 02 14:32:04 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg509his' Tue Jun 30 17:01:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg509His' P15056:p.Arg662Leu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R L Wed May 02 14:57:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg662leu' Tue Jun 30 18:08:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg662Leu' 6 unspecified role unspecified role bait bait Q02750:p.Gln56Pro mutation mutation certain certain sequence position certain certain certain sequence position certain Q P Wed May 02 16:38:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln56pro' Tue Jun 30 15:58:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln56Pro' 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 physical association physical association 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 4 7 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 P15056:p.Val600Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain V E Wed May 02 14:36:02 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 19:39:22 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' 6 unspecified role unspecified role bait bait Q02750:p.Glu203Lys mutation mutation certain certain sequence position certain certain certain sequence position certain E K Wed May 02 20:11:23 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu203lys' Tue Jun 30 19:46:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu203Lys' 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 physical association physical association Supp 4 map2k1-braf-21 4 7 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 P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain V E Wed May 02 18:35:06 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 19:09:34 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' 6 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 K . Wed May 02 15:56:54 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys59del' Tue Jun 30 17:00:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys59del' 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 physical association physical association Supp 4i An in-frame deletion mutant (K59) showed binding to the BRAF V600E mutant but not the WT. Is there any other way to show this? I cannot enter surrounding sequences to K59 to show the deletion as I intended. map2k1-braf-22 5 7 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 P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain V E Wed May 02 17:23:02 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 17:05:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' 6 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 K . Wed May 02 16:19:45 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys59del' Tue Jun 30 18:31:46 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys59del' 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 physical association physical association Supp 4i An in-frame deletion mutant (K59) showed binding to the BRAF V600E mutant but not the WT. Is there any other way to show this? I cannot enter surrounding sequences to K59 to show the deletion as I intended. map2k1-braf-23 5 7 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 P15056:p.Val600Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain V E Wed May 02 15:18:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 18:43:39 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' P15056:p.Arg509His mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R H Wed May 02 14:53:40 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg509his' Tue Jun 30 19:41:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg509His' P15056:p.Arg662Leu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R L Wed May 02 14:41:17 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg662leu' Tue Jun 30 19:25:53 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg662Leu' 6 unspecified role unspecified role bait bait Q02750:p.Gln56Pro mutation mutation certain certain sequence position certain certain certain sequence position certain Q P Wed May 02 14:34:54 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln56pro' Tue Jun 30 19:22:48 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln56Pro' 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 physical association physical association 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 5 7 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 P15056:p.Arg509His mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R H Wed May 02 19:45:17 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg509his' Tue Jun 30 16:17:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg509His' P15056:p.Arg662Leu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R L Wed May 02 16:26:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg662leu' Tue Jun 30 19:18:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg662Leu' P15056:p.Val600Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain V E Wed May 02 16:51:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 16:35:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' 6 unspecified role unspecified role bait bait Q02750:p.Phe129Leu mutation mutation certain certain sequence position certain certain certain sequence position certain F L Wed May 02 19:23:17 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe129leu' Tue Jun 30 16:18:19 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe129Leu' 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 physical association physical association 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 5 7 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 P15056:p.Val600Glu mutation mutation certain certain sequence position certain certain certain sequence position certain V E Wed May 02 14:43:05 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val600glu' Tue Jun 30 18:17:57 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val600Glu' 6 unspecified role unspecified role bait bait Q02750:p.Glu203Lys mutation mutation certain certain sequence position certain certain certain sequence position certain E K Wed May 02 20:14:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu203lys' Tue Jun 30 19:18:38 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu203Lys' 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 physical association physical association Supp 4