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/ Disruption of CRAF-Mediated MEK Activation Is Required for Effective MEK Inhibition in KRAS Mutant Tumors. Disruption of CRAF-Mediated MEK Activation Is Required for Effective MEK Inhibition in KRAS Mutant Tumors. Cancer Cell (1535-6108) 2014 imex curation Lito P., Saborowski A., Yue J., Solomon M., Joseph E., Gadal S., Saborowski M., Kastenhuber E., Fellmann C., Ohara K., Morikami K., Miura T., Lukacs C., Ishii N., Lowe S., Rosen N. Cancer - Interactions investigated in the context of cancer MEK1-, KRas-, and NRas-encoding plasmids are stated to have been obtained from Biomyx; constructs containing these proteins have been entered as Human, as no equivalent mouse products seem to be available on the Biomyx website. rosenn@mskcc.org,lowes@mskcc.org Only protein-protein interactions human-a_549 Homo sapiens Caucasian lung carcinoma cell line a_549 Human Caucasian lung carcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Time of PD0325901 treatment (hours) 0 3 48 Time of Trametinib treatment (hours) 48 0 3 Lito P., Saborowski A., Yue J., Solomon M., Joseph E., Gadal S., Saborowski M., Kastenhuber E., Fellmann C., Ohara K., Morikami K., Miura T., Lukacs C., Ishii N., Lowe S., Rosen N. Cancer - Interactions investigated in the context of cancer MEK1-, KRas-, and NRas-encoding plasmids are stated to have been obtained from Biomyx; constructs containing these proteins have been entered as Human, as no equivalent mouse products seem to be available on the Biomyx website. rosenn@mskcc.org,lowes@mskcc.org Accepted 2015-MAY-27 by PPORRAS Time of drug treatment Drug used Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation 6D added as a separate Expt as there are variable conditions, and the interaction has one less interactor than 6C.; I have - eventually- managed to do as you suggest - not easy as if you make a single mistake and press "save" everything is lost....(cant remember exactly what happens - but it means everything has to be correct the first time of entry, which when people are talking across you is not easy...) Disruption of CRAF-Mediated MEK Activation Is Required for Effective MEK Inhibition in KRAS Mutant Tumors. Disruption of CRAF-Mediated MEK Activation Is Required for Effective MEK Inhibition in KRAS Mutant Tumors. Cancer Cell (1535-6108) 2014 imex curation Lito P., Saborowski A., Yue J., Solomon M., Joseph E., Gadal S., Saborowski M., Kastenhuber E., Fellmann C., Ohara K., Morikami K., Miura T., Lukacs C., Ishii N., Lowe S., Rosen N. Cancer - Interactions investigated in the context of cancer MEK1-, KRas-, and NRas-encoding plasmids are stated to have been obtained from Biomyx; constructs containing these proteins have been entered as Human, as no equivalent mouse products seem to be available on the Biomyx website. rosenn@mskcc.org,lowes@mskcc.org Only protein-protein interactions in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Kd - no drug treatment molar molar M M 5.51E-08 3.83E-08 Kd PD0325901 treatment (unknown concentration) molar molar M M 4.99E-08 1.66E-08 Kd Trametinib treatment - 3uM molar molar M M 1.01E-07 1.07E-07 Kd CH5126766 treatment (unknown concentration) molar molar M M 2.30E-09 6.78E-10 Lito P., Saborowski A., Yue J., Solomon M., Joseph E., Gadal S., Saborowski M., Kastenhuber E., Fellmann C., Ohara K., Morikami K., Miura T., Lukacs C., Ishii N., Lowe S., Rosen N. Cancer - Interactions investigated in the context of cancer MEK1-, KRas-, and NRas-encoding plasmids are stated to have been obtained from Biomyx; constructs containing these proteins have been entered as Human, as no equivalent mouse products seem to be available on the Biomyx website. rosenn@mskcc.org,lowes@mskcc.org Accepted 2015-MAY-27 by PPORRAS Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation 6D added as a separate Expt as there are variable conditions, and the interaction has one less interactor than 6C.; I have - eventually- managed to do as you suggest - not easy as if you make a single mistake and press "save" everything is lost....(cant remember exactly what happens - but it means everything has to be correct the first time of entry, which when people are talking across you is not easy...) Disruption of CRAF-Mediated MEK Activation Is Required for Effective MEK Inhibition in KRAS Mutant Tumors. Disruption of CRAF-Mediated MEK Activation Is Required for Effective MEK Inhibition in KRAS Mutant Tumors. Cancer Cell (1535-6108) 2014 imex curation Lito P., Saborowski A., Yue J., Solomon M., Joseph E., Gadal S., Saborowski M., Kastenhuber E., Fellmann C., Ohara K., Morikami K., Miura T., Lukacs C., Ishii N., Lowe S., Rosen N. Cancer - Interactions investigated in the context of cancer MEK1-, KRas-, and NRas-encoding plasmids are stated to have been obtained from Biomyx; constructs containing these proteins have been entered as Human, as no equivalent mouse products seem to be available on the Biomyx website. rosenn@mskcc.org,lowes@mskcc.org Only protein-protein interactions in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Lito P., Saborowski A., Yue J., Solomon M., Joseph E., Gadal S., Saborowski M., Kastenhuber E., Fellmann C., Ohara K., Morikami K., Miura T., Lukacs C., Ishii N., Lowe S., Rosen N. Cancer - Interactions investigated in the context of cancer MEK1-, KRas-, and NRas-encoding plasmids are stated to have been obtained from Biomyx; constructs containing these proteins have been entered as Human, as no equivalent mouse products seem to be available on the Biomyx website. rosenn@mskcc.org,lowes@mskcc.org Accepted 2015-MAY-27 by PPORRAS Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation 6D added as a separate Expt as there are variable conditions, and the interaction has one less interactor than 6C.; I have - eventually- managed to do as you suggest - not easy as if you make a single mistake and press "save" everything is lost....(cant remember exactly what happens - but it means everything has to be correct the first time of entry, which when people are talking across you is not easy...) Disruption of CRAF-Mediated MEK Activation Is Required for Effective MEK Inhibition in KRAS Mutant Tumors. Disruption of CRAF-Mediated MEK Activation Is Required for Effective MEK Inhibition in KRAS Mutant Tumors. Cancer Cell (1535-6108) 2014 imex curation Lito P., Saborowski A., Yue J., Solomon M., Joseph E., Gadal S., Saborowski M., Kastenhuber E., Fellmann C., Ohara K., Morikami K., Miura T., Lukacs C., Ishii N., Lowe S., Rosen N. Cancer - Interactions investigated in the context of cancer MEK1-, KRas-, and NRas-encoding plasmids are stated to have been obtained from Biomyx; constructs containing these proteins have been entered as Human, as no equivalent mouse products seem to be available on the Biomyx website. rosenn@mskcc.org,lowes@mskcc.org Only protein-protein interactions human-a375 Human malignant melanoma a375 Human malignant melanoma ATCC catalog # CRL-1872 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Lito P., Saborowski A., Yue J., Solomon M., Joseph E., Gadal S., Saborowski M., Kastenhuber E., Fellmann C., Ohara K., Morikami K., Miura T., Lukacs C., Ishii N., Lowe S., Rosen N. Cancer - Interactions investigated in the context of cancer MEK1-, KRas-, and NRas-encoding plasmids are stated to have been obtained from Biomyx; constructs containing these proteins have been entered as Human, as no equivalent mouse products seem to be available on the Biomyx website. rosenn@mskcc.org,lowes@mskcc.org Accepted 2015-MAY-27 by PPORRAS Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation 6D added as a separate Expt as there are variable conditions, and the interaction has one less interactor than 6C.; I have - eventually- managed to do as you suggest - not easy as if you make a single mistake and press "save" everything is lost....(cant remember exactly what happens - but it means everything has to be correct the first time of entry, which when people are talking across you is not easy...) Disruption of CRAF-Mediated MEK Activation Is Required for Effective MEK Inhibition in KRAS Mutant Tumors. Disruption of CRAF-Mediated MEK Activation Is Required for Effective MEK Inhibition in KRAS Mutant Tumors. Cancer Cell (1535-6108) 2014 imex curation Lito P., Saborowski A., Yue J., Solomon M., Joseph E., Gadal S., Saborowski M., Kastenhuber E., Fellmann C., Ohara K., Morikami K., Miura T., Lukacs C., Ishii N., Lowe S., Rosen N. Cancer - Interactions investigated in the context of cancer MEK1-, KRas-, and NRas-encoding plasmids are stated to have been obtained from Biomyx; constructs containing these proteins have been entered as Human, as no equivalent mouse products seem to be available on the Biomyx website. rosenn@mskcc.org,lowes@mskcc.org Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Lito P., Saborowski A., Yue J., Solomon M., Joseph E., Gadal S., Saborowski M., Kastenhuber E., Fellmann C., Ohara K., Morikami K., Miura T., Lukacs C., Ishii N., Lowe S., Rosen N. Cancer - Interactions investigated in the context of cancer MEK1-, KRas-, and NRas-encoding plasmids are stated to have been obtained from Biomyx; constructs containing these proteins have been entered as Human, as no equivalent mouse products seem to be available on the Biomyx website. rosenn@mskcc.org,lowes@mskcc.org Accepted 2015-MAY-27 by PPORRAS Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation 6D added as a separate Expt as there are variable conditions, and the interaction has one less interactor than 6C.; I have - eventually- managed to do as you suggest - not easy as if you make a single mistake and press "save" everything is lost....(cant remember exactly what happens - but it means everything has to be correct the first time of entry, which when people are talking across you is not easy...) Disruption of CRAF-Mediated MEK Activation Is Required for Effective MEK Inhibition in KRAS Mutant Tumors. Disruption of CRAF-Mediated MEK Activation Is Required for Effective MEK Inhibition in KRAS Mutant Tumors. Cancer Cell (1535-6108) 2014 imex curation Lito P., Saborowski A., Yue J., Solomon M., Joseph E., Gadal S., Saborowski M., Kastenhuber E., Fellmann C., Ohara K., Morikami K., Miura T., Lukacs C., Ishii N., Lowe S., Rosen N. Cancer - Interactions investigated in the context of cancer MEK1-, KRas-, and NRas-encoding plasmids are stated to have been obtained from Biomyx; constructs containing these proteins have been entered as Human, as no equivalent mouse products seem to be available on the Biomyx website. rosenn@mskcc.org,lowes@mskcc.org Only protein-protein interactions human-a_549 Homo sapiens Caucasian lung carcinoma cell line a_549 Human Caucasian lung carcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Time of PD0325901 treatment (hours) 0 3 48 Time of Trametinib treatment (hours) 48 0 3 Lito P., Saborowski A., Yue J., Solomon M., Joseph E., Gadal S., Saborowski M., Kastenhuber E., Fellmann C., Ohara K., Morikami K., Miura T., Lukacs C., Ishii N., Lowe S., Rosen N. Cancer - Interactions investigated in the context of cancer I have added BOTH ways of adding Variable conditions as you cannot see from the summary view what you have added to which interaction. (Variable condits have to be in a separate Expt) MEK1-, KRas-, and NRas-encoding plasmids are stated to have been obtained from Biomyx; constructs containing these proteins have been entered as Human, as no equivalent mouse products seem to be available on the Biomyx website. rosenn@mskcc.org,lowes@mskcc.org Accepted 2015-MAY-27 by PPORRAS Time of drug treatment Drug used Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation 6D added as a separate Expt as there are variable conditions, and the interaction has one less interactor than 6C.; I have - eventually- managed to do as you suggest - not easy as if you make a single mistake and press "save" everything is lost....(cant remember exactly what happens - but it means everything has to be correct the first time of entry, which when people are talking across you is not easy...) 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 araf_human Serine/threonine-protein kinase A-Raf ARAF PKS PKS2 ARAF1 Proto-oncogene A-Raf-1 Proto-oncogene Pks Proto-oncogene A-Raf protein protein human Homo sapiens Human MEPPRGPPANGAEPSRAVGTVKVYLPNKQRTVVTVRDGMSVYDSLDKALKVRGLNQDCCVVYRLIKGRKTVTAWDTAIAPLDGEELIVEVLEDVPLTMHNFVRKTFFSLAFCDFCLKFLFHGFRCQTCGYKFHQHCSSKVPTVCVDMSTNRQQFYHSVQDLSGGSRQHEAPSNRPLNELLTPQGPSPRTQHCDPEHFPFPAPANAPLQRIRSTSTPNVHMVSTTAPMDSNLIQLTGQSFSTDAAGSRGGSDGTPRGSPSPASVSSGRKSPHSKSPAEQRERKSLADDKKKVKNLGYRDSGYYWEVPPSEVQLLKRIGTGSFGTVFRGRWHGDVAVKVLKVSQPTAEQAQAFKNEMQVLRKTRHVNILLFMGFMTRPGFAIITQWCEGSSLYHHLHVADTRFDMVQLIDVARQTAQGMDYLHAKNIIHRDLKSNNIFLHEGLTVKIGDFGLATVKTRWSGAQPLEQPSGSVLWMAAEVIRMQDPNPYSFQSDVYAYGVVLYELMTGSLPYSHIGCRDQIIFMVGRGYLSPDLSKISSNCPKAMRRLLSDCLKFQREERPLFPQILATIELLQRSLPKIERSASEPSLHRTQADELPACLLSAARLVP DIP protein P10398 original sequence version: 126 DIP protein P10398 original sequence version: 168 D23E5711304AA468 p04049-1 RAF proto-oncogene serine/threonine-protein kinase 6C RAF1 RAF Proto-oncogene c-RAF Raf-1 protein protein human Homo sapiens Human MEHIQGAWKTISNGFGFKDAVFDGSSCISPTIVQQFGYQRRASDDGKLTDPSKTSNTIRVFLPNKQRTVVNVRNGMSLHDCLMKALKVRGLQPECCAVFRLLHEHKGKKARLDWNTDAASLIGEELQVDFLDHVPLTTHNFARKTFLKLAFCDICQKFLLNGFRCQTCGYKFHEHCSTKVPTMCVDWSNIRQLLLFPNSTIGDSGVPALPSLTMRRMRESVSRMPVSSQHRYSTPHAFTFNTSSPSSEGSLSQRQRSTSTPNVHMVSTTLPVDSRMIEDAIRSHSESASPSALSSSPNNLSPTGWSQPKTPVPAQRERAPVSGTQEKNKIRPRGQRDSSYYWEIEASEVMLSTRIGSGSFGTVYKGKWHGDVAVKILKVVDPTPEQFQAFRNEVAVLRKTRHVNILLFMGYMTKDNLAIVTQWCEGSSLYKHLHVQETKFQMFQLIDIARQTAQGMDYLHAKNIIHRDMKSNNIFLHEGLTVKIGDFGLATVKSRWSGSQQVEQPTGSVLWMAPEVIRMQDNNPFSFQSDVYSYGIVLYELMTGELPYSHINNRDQIIFMVGRGYASPDLSKLYKNCPKAMKRLVADCVKKVKEERPLFPQILSSIELLQHSLPKINRSASEPSLHRAAHTEDINACTLTTSPRLPVF EF821B5349711BC3 ch5126766 CH5126766 small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) 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 raf1_human RAF proto-oncogene serine/threonine-protein kinase RAF1 RAF Proto-oncogene c-RAF Raf-1 protein protein human Homo sapiens Human MEHIQGAWKTISNGFGFKDAVFDGSSCISPTIVQQFGYQRRASDDGKLTDPSKTSNTIRVFLPNKQRTVVNVRNGMSLHDCLMKALKVRGLQPECCAVFRLLHEHKGKKARLDWNTDAASLIGEELQVDFLDHVPLTTHNFARKTFLKLAFCDICQKFLLNGFRCQTCGYKFHEHCSTKVPTMCVDWSNIRQLLLFPNSTIGDSGVPALPSLTMRRMRESVSRMPVSSQHRYSTPHAFTFNTSSPSSEGSLSQRQRSTSTPNVHMVSTTLPVDSRMIEDAIRSHSESASPSALSSSPNNLSPTGWSQPKTPVPAQRERAPVSGTQEKNKIRPRGQRDSSYYWEIEASEVMLSTRIGSGSFGTVYKGKWHGDVAVKILKVVDPTPEQFQAFRNEVAVLRKTRHVNILLFMGYMTKDNLAIVTQWCEGSSLYKHLHVQETKFQMFQLIDIARQTAQGMDYLHAKNIIHRDMKSNNIFLHEGLTVKIGDFGLATVKSRWSGSQQVEQPTGSVLWMAPEVIRMQDNNPFSFQSDVYSYGIVLYELMTGELPYSHINNRDQIIFMVGRGYASPDLSKLYKNCPKAMKRLVADCVKKVKEERPLFPQILSSIELLQHSLPKINRSASEPSLHRAAHTEDINACTLTTSPRLPVF DIP protein P04049 original sequence version: 117 DIP protein P04049 original sequence version: 106 DIP protein P04049 original sequence version: 114 DIP protein P04049 original sequence version: 141 DIP protein P04049 original sequence version: 142 DIP protein P04049 original sequence version: 147 DIP protein P04049-1 original sequence version: 136 DIP protein P04049 original sequence version: 136 DIP protein P04049 original sequence version: 153 DIP protein P04049 original sequence version: 150 DIP protein P04049 original sequence version: 164 DIP protein P04049 original sequence version: 172 DIP protein P04049-1 original sequence version: 172 DIP protein P04049 original sequence version: 184 DIP protein P04049 original sequence version: 188 DIP protein P04049 original sequence version: 195 EF821B5349711BC3 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 phosphoaminophosphonic acid-adenylate ester PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) map2k1-araf-4 19 28 unspecified role unspecified role prey prey 32 unspecified role unspecified role prey prey 33 unspecified role unspecified role prey prey 31 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 Q02750:p.[Ser218Glu;Ser222Glu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain S E certain certain sequence position certain certain certain sequence position certain S E Wed May 02 18:06:03 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser218glu,ser222glu' Tue Jun 30 17:10:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser218Glu;Ser222Glu]' Q02750:p.[Met308Ala;Ile310Ala] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain M A certain certain sequence position certain certain certain sequence position certain I A Wed May 02 17:58:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'met308ala,ile310ala' Tue Jun 30 16:00:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Met308Ala;Ile310Ala]' association association 4 A/D, 6A, 7G, Supp figure 4 A MEK inhibitors: Selumetinib and PD0325901 CH5126766 - a newer MEK inhibitor. Trametinib -a newer MEK inhibitor, disrupts the interaction of MEK1 with ARAF and CRAF. In cells co-transfected with K-RAS G12V this interaction was completely disrupted, but was only partially reduced in the presence of CH5126766 or PD0325901. Co-transfection of BRAF V600E reduced this interaction,and the presence of CH5126766 or PD0325901 further reduced the interaction , but in the presence of CH5126766 the interaction was increased. map2k1-araf-1 18 31 unspecified role unspecified role bait bait 33 unspecified role unspecified role prey prey 28 unspecified role unspecified role prey prey 32 unspecified role unspecified role prey prey association association 4 B/C, 6B The MEK inhibitor PD0325901 reduced the level of MEK1 in complex with BRAF and ARAF. map2k1-araf-2 1 33 unspecified role unspecified role prey prey 31 unspecified role unspecified role bait bait 32 unspecified role unspecified role prey prey 28 unspecified role unspecified role prey prey association association 3 4 C, 6C The MEK inhibitor PD0325901 Trametinib - a newer MEK inhibitor braf-map2k1-1 8 31 unspecified role unspecified role unspecified role unspecified role n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 33 unspecified role unspecified role unspecified role unspecified role 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 kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction 9 15 13 11 6 E/F Trametinib is a MEK inhibitor that reduces the affinity of MEK and RAF. raf1-map2k1-2 8 31 unspecified role unspecified role prey prey n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 29 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 kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P04049-1:p.Tyr340_Tyr341delinsAspAsp mutation mutation certain certain sequence position certain certain certain sequence position certain YY DD Thu Jul 07 12:16:49 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was tyr340asp,tyr341asp Fri Jul 22 11:00:15 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr340asp,tyr341asp' Wed May 02 15:49:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr_tyr340-341asp_asp' Tue Jun 30 19:59:19 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr340_Tyr341delinsAspAsp' direct interaction direct interaction 12 16 14 10 6 E/F Trametinib is a MEK inhibitor that reduces the affinity of MEK and RAF. ch5126766-map2k1-1 17 30 allosteric effector allosteric effector neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 34 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 31 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 regions sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Table S1 2.7 A R-factor: working - 21.7%, Free R-factor: 25.0%, map2k2-araf-4 19 28 unspecified role unspecified role prey prey 27 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 33 unspecified role unspecified role prey prey 32 unspecified role unspecified role prey prey association association CH5126766 - a newer MEK inhibitor. (PD0325901) K-RAS G12V transfection:- this interaction was completely disrupted, but was only partially reduced in the presence of CH5126766 or PD0325901. BRAF V600E transfection: MEK2 interacted with ARAF, BRAF and CRAF, but no untransfected control result is shown. The presence of PD0325901 did not appear to affect this interaction. In the presence of Trametinib the interaction of MEK2 and BRAF still took place at a reduced level. It is not clear if any of these interactions were with endogenous wild-type BRAF, or the V600E mutant, or both. Supp fig 4B Trametinib -a newer MEK inhibitor, disrupts the interaction of MEK2 with ARAF and CRAF. map2k1-araf-6 20 32 unspecified role unspecified role prey prey 33 unspecified role unspecified role prey prey 31 unspecified role unspecified role bait bait association association 4 6 6D The MEK inhibitor PD0325901 Trametinib - a newer MEK inhibitor map2k1-braf-26 20 33 unspecified role unspecified role prey prey 31 unspecified role unspecified role bait bait physical association physical association 2 6D The MEK inhibitor PD0325901 Trametinib - a newer MEK inhibitor weak interaction 0 hours PD0325901 map2k1-araf-7 20 33 unspecified role unspecified role prey prey 31 unspecified role unspecified role bait bait 32 unspecified role unspecified role prey prey association association 26 6D The MEK inhibitor PD0325901 Trametinib - a newer MEK inhibitor weak interaction Time of Trametinib treatment (hours)-3 map2k1-braf-27 20 31 unspecified role unspecified role bait bait 32 unspecified role unspecified role prey prey physical association physical association 24 6D The MEK inhibitor PD0325901 Trametinib - a newer MEK inhibitor weak interaction 48 hours Trametinib