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/ levinson-2008-1 Structural basis for the recognition of c-Src by its inactivator Csk. in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr southern blot southern blot Levinson NM., Seeliger MA., Cole PA., Kuriyan J. kuriyan@berkeley.edu Cell (0092-8674) 2008 Only protein-protein interactions 5 imex curation levinson-2008-2 Structural basis for the recognition of c-Src by its inactivator Csk. in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Levinson NM., Seeliger MA., Cole PA., Kuriyan J. kuriyan@berkeley.edu Cell (0092-8674) 2008 Only protein-protein interactions 5 imex curation levinson-2008-3 Structural basis for the recognition of c-Src by its inactivator Csk. in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Levinson NM., Seeliger MA., Cole PA., Kuriyan J. kuriyan@berkeley.edu Cell (0092-8674) 2008 Only protein-protein interactions 5 imex curation src_chick Proto-oncogene tyrosine-protein kinase Src SRC pp60c-src Proto-oncogene c-Src protein protein chick Gallus gallus (Chicken) Chicken MGSSKSKPKDPSQRRRSLEPPDSTHHGGFPASQTPNKTAAPDTHRTPSRSFGTVATEPKLFGGFNTSDTVTSPQRAGALAGGVTTFVALYDYESRTETDLSFKKGERLQIVNNTEGDWWLAHSLTTGQTGYIPSNYVAPSDSIQAEEWYFGKITRRESERLLLNPENPRGTFLVRESETTKGAYCLSVSDFDNAKGLNVKHYKIRKLDSGGFYITSRTQFSSLQQLVAYYSKHADGLCHRLTNVCPTSKPQTQGLAKDAWEIPRESLRLEVKLGQGCFGEVWMGTWNGTTRVAIKTLKPGTMSPEAFLQEAQVMKKLRHEKLVQLYAVVSEEPIYIVTEYMSKGSLLDFLKGEMGKYLRLPQLVDMAAQIASGMAYVERMNYVHRDLRAANILVGENLVCKVADFGLARLIEDNEYTARQGAKFPIKWTAPEAALYGRFTIKSDVWSFGILLTELTTKGRVPYPGMVNREVLDQVERGYRMPCPPECPESLHDLMCQCWRKDPEERPTFEYLQAFLEDYFTSTEPQYQPGENL csk_human Tyrosine-protein kinase CSK CSK C-Src kinase Protein-tyrosine kinase CYL protein protein human Homo sapiens Human MSAIQAAWPSGTECIAKYNFHGTAEQDLPFCKGDVLTIVAVTKDPNWYKAKNKVGREGIIPANYVQKREGVKAGTKLSLMPWFHGKITREQAERLLYPPETGLFLVRESTNYPGDYTLCVSCDGKVEHYRIMYHASKLSIDEEVYFENLMQLVEHYTSDADGLCTRLIKPKVMEGTVAAQDEFYRSGWALNMKELKLLQTIGKGEFGDVMLGDYRGNKVAVKCIKNDATAQAFLAEASVMTQLRHSNLVQLLGVIVEEKGGLYIVTEYMAKGSLVDYLRSRGRSVLGGDCLLKFSLDVCEAMEYLEGNNFVHRDLAARNVLVSEDNVAKVSDFGLTKEASSTQDTGKLPVKWTAPEALREKKFSTKSDVWSFGILLWEIYSFGRVPYPRIPLKDVVPRVEKGYKMDAPDGCPPAVYEVMKNCWHLDAAMRPSFLQLREQLEHIKTHELHL staurosporine Staurosporine small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) src-csk-1 Interaction between chick src and human csk demonstrated by SPR 760903 N/A 760906 unspecified role unspecified role bait bait kinase region 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) N/A 760907 unspecified role unspecified role prey prey kinase region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) 760913 760911 direct interaction direct interaction false false false Full length Csk vs kinase region of Src KD=2.6E-5M SH2 + kinase region (aa 68-450) Csk vs kinase region of Src KD=3.1E-5M Full length Csk vs SH2 + SH3 + kinase region of Src (aa 83-533)KD=2.0E-5M Fig. 1, Supp Fig. 1 Interaction is dependent on salt concentration, with higher salt concentrations weakening binding. src-csk-4 Interaction between chick src and human csk demonstrated by SPR 760903 N/A-x 760907 unspecified role unspecified role prey prey P41240:p.Arg279Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P41240:p.Arg281Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P41240:p.Arg283Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P41240:p.Arg384Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P41240:p.Arg389Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) N/A-x 760906 unspecified role unspecified role bait bait kinase region 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) direct interaction direct interaction false false false Full length Csk vs kinase region of Src KD=2.6E-5M Fig. 1, Fig. 3D Interaction is dependent on salt concentration, higher salt concentrations weakens binding. src-csk-5 Interaction between chick src and human csk demonstrated by SPR 760903 N/A 760906 unspecified role unspecified role bait bait P00523:p.Lys442Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false kinase region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P00523:p.Asp518Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 760907 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false Full length Csk vs kinase region of Src KD=2.6E-5M Fig. 1, Fig. 3D Interaction is dependent on salt concentration, higher salt concentrations weakens binding. src-csk-2 Interaction between chick src, human csk and staurosporine demonstrated by X-ray crystallography 760904 N/A 760906 unspecified role unspecified role neutral component neutral component kinase region 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) N/A 760908 unspecified role unspecified role neutral component neutral component N/A 760907 unspecified role unspecified role neutral component neutral component kinase region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false Fig. 2, Supp. Table 1 Working 23.7%, free 27.8% 2.9A src-csk-3 Interaction between chick src and human csk demonstrated by X-ray crystallography 760904 N/A-x-x 760906 unspecified role unspecified role neutral component neutral component sh2_sh3_kin_region 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) N/A-x-x 760907 unspecified role unspecified role neutral component neutral component kinase region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) 760939 760937 direct interaction direct interaction false false false Fig. 2, Supp. Table 1 Working 29.4%, free 30.2% 4.11A src-csk-6 Phosphorylation of chick src by human csk 760905 N/A-x-x 760907 enzyme enzyme neutral component neutral component ecobb Escherichia coli (strain B / BL21) N/A-x-x 760906 enzyme target enzyme target substrate substrate neutral component neutral component P00523:p.Glu510Ala Alters kinetics of interaction mutation mutation certain certain sequence position certain certain certain sequence position certain false P00523:p.Lys442Ala Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false kinase region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P00523:p.Asp518Ala Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P00523:p.Lys295Met mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false Fig. 3E The lys295met mutant prevented autophosphorylation Phosphorylation of Src