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/ p27 phosphorylation by Src regulates inhibition of cyclin E-Cdk2. p27 phosphorylation by Src regulates inhibition of cyclin E-Cdk2. Cell (0092-8674) 2007 imex curation Chu I., Sun J., Arnaout A., Kahn H., Hanna W., Narod S., Sun P., Tan CK., Hengst L., Slingerland J. Cancer - Interactions investigated in the context of cancer CDN1B/p27 identified as of human origin in Grimmler et al. 2007 (PMID: 17254966). Details of c-Src taken from manufacturer's website. jslingerland@med.miami.edu 02-MAR-2006: Contacted by JYOTI. Only protein-protein interactions 8 in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Chu I., Sun J., Arnaout A., Kahn H., Hanna W., Narod S., Sun P., Tan CK., Hengst L., Slingerland J. Cancer - Interactions investigated in the context of cancer CDN1B/p27 identified as of human origin in Grimmler et al. 2007 (PMID: 17254966). Details of c-Src taken from manufacturer's website. jslingerland@med.miami.edu Accepted 2007-FEB-15 by DAVET. 02-MAR-2006: Contacted by JYOTI. Cell (0092-8674) 2007 Only protein-protein interactions 8 imex curation p27 phosphorylation by Src regulates inhibition of cyclin E-Cdk2. p27 phosphorylation by Src regulates inhibition of cyclin E-Cdk2. Cell (0092-8674) 2007 imex curation Chu I., Sun J., Arnaout A., Kahn H., Hanna W., Narod S., Sun P., Tan CK., Hengst L., Slingerland J. Cancer - Interactions investigated in the context of cancer CDN1B/p27 identified as of human origin in Grimmler et al. 2007 (PMID: 17254966). Details of c-Src taken from manufacturer's website. jslingerland@med.miami.edu 02-MAR-2006: Contacted by JYOTI. Only protein-protein interactions 8 in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Chu I., Sun J., Arnaout A., Kahn H., Hanna W., Narod S., Sun P., Tan CK., Hengst L., Slingerland J. Cancer - Interactions investigated in the context of cancer jslingerland@med.miami.edu Accepted 2007-FEB-15 by DAVET. 02-MAR-2006: Contacted by JYOTI. Cell (0092-8674) 2007 Only protein-protein interactions 8 imex curation p27 phosphorylation by Src regulates inhibition of cyclin E-Cdk2. p27 phosphorylation by Src regulates inhibition of cyclin E-Cdk2. Cell (0092-8674) 2007 imex curation Chu I., Sun J., Arnaout A., Kahn H., Hanna W., Narod S., Sun P., Tan CK., Hengst L., Slingerland J. Cancer - Interactions investigated in the context of cancer CDN1B/p27 identified as of human origin in Grimmler et al. 2007 (PMID: 17254966). Details of c-Src taken from manufacturer's website. jslingerland@med.miami.edu 02-MAR-2006: Contacted by JYOTI. Only protein-protein interactions 8 human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Chu I., Sun J., Arnaout A., Kahn H., Hanna W., Narod S., Sun P., Tan CK., Hengst L., Slingerland J. Cancer - Interactions investigated in the context of cancer jslingerland@med.miami.edu Accepted 2007-FEB-15 by DAVET. 02-MAR-2006: Contacted by JYOTI. Cell (0092-8674) 2007 Only protein-protein interactions 8 imex curation yes_human Tyrosine-protein kinase Yes p61-Yes Proto-oncogene c-Yes YES1 YES protein protein human Homo sapiens Human MGCIKSKENKSPAIKYRPENTPEPVSTSVSHYGAEPTTVSPCPSSSAKGTAVNFSSLSMTPFGGSSGVTPFGGASSSFSVVPSSYPAGLTGGVTIFVALYDYEARTTEDLSFKKGERFQIINNTEGDWWEARSIATGKNGYIPSNYVAPADSIQAEEWYFGKMGRKDAERLLLNPGNQRGIFLVRESETTKGAYSLSIRDWDEIRGDNVKHYKIRKLDNGGYYITTRAQFDTLQKLVKHYTEHADGLCHKLTTVCPTVKPQTQGLAKDAWEIPRESLRLEVKLGQGCFGEVWMGTWNGTTKVAIKTLKPGTMMPEAFLQEAQIMKKLRHDKLVPLYAVVSEEPIYIVTEFMSKGSLLDFLKEGDGKYLKLPQLVDMAAQIADGMAYIERMNYIHRDLRAANILVGENLVCKIADFGLARLIEDNEYTARQGAKFPIKWTAPEAALYGRFTIKSDVWSFGILQTELVTKGRVPYPGMVNREVLEQVERGYRMPCPQGCPESLHELMNLCWKKDPDERPTFEYIQSFLEDYFTATEPQYQPGENL DIP protein P07947 original sequence version: 131 DIP protein P07947 original sequence version: 148 DIP protein P07947 original sequence version: 140 A2B376084686BBCD src_human Proto-oncogene tyrosine-protein kinase Src pp60c-src SRC SRC1 Proto-oncogene c-Src protein protein human Homo sapiens Human MGSNKSKPKDASQRRRSLEPAENVHGAGGGAFPASQTPSKPASADGHRGPSAAFAPAAAEPKLFGGFNSSDTVTSPQRAGPLAGGVTTFVALYDYESRTETDLSFKKGERLQIVNNTEGDWWLAHSLSTGQTGYIPSNYVAPSDSIQAEEWYFGKITRRESERLLLNAENPRGTFLVRESETTKGAYCLSVSDFDNAKGLNVKHYKIRKLDSGGFYITSRTQFNSLQQLVAYYSKHADGLCHRLTTVCPTSKPQTQGLAKDAWEIPRESLRLEVKLGQGCFGEVWMGTWNGTTRVAIKTLKPGTMSPEAFLQEAQVMKKLRHEKLVQLYAVVSEEPIYIVTEYMSKGSLLDFLKGETGKYLRLPQLVDMAAQIASGMAYVERMNYVHRDLRAANILVGENLVCKVADFGLARLIEDNEYTARQGAKFPIKWTAPEAALYGRFTIKSDVWSFGILLTELTTKGRVPYPGMVNREVLDQVERGYRMPCPPECPESLHDLMCQCWRKEPEERPTFEYLQAFLEDYFTSTEPQYQPGENL DIP protein P12931 original sequence version: 188 DIP protein P12931 original sequence version: 189 DIP protein P12931 original sequence version: 190 DIP protein P12931 original sequence version: 191 DIP protein P12931 original sequence version: 115 DIP protein P12931 original sequence version: 116 DIP protein P12931 original sequence version: 164 DIP protein P12931 original sequence version: 142 DIP protein P12931 original sequence version: 143 DIP protein P12931 original sequence version: 150 DIP protein P12931 original sequence version: 160 DIP protein P12931 original sequence version: 184 DIP protein P12931 original sequence version: 155 DIP protein P12931-1 original sequence version: 155 DIP protein P12931 original sequence version: 202 C1908084683E5DE8 cdn1b_human Cyclin-dependent kinase inhibitor 1B Cyclin-dependent kinase inhibitor p27 p27Kip1 CDKN1B KIP1 protein protein human Homo sapiens Human MSNVRVSNGSPSLERMDARQAEHPKPSACRNLFGPVDHEELTRDLEKHCRDMEEASQRKWNFDFQNHKPLEGKYEWQEVEKGSLPEFYYRPPRPPKGACKVPAQESQDVSGSRPAAPLIGAPANSEDTHLVDPKTDPSDSQTGLAEQCAGIRKRPATDDSSTQNKRANRTEENVSDGSPNAGSVEQTPKKPGLRRRQT DIP protein P46527 original sequence version: 120 DIP protein P46527 original sequence version: 115 DIP protein P46527 original sequence version: 154 DIP protein P46527 original sequence version: 168 DIP protein P46527 original sequence version: 164 DIP protein P46527 original sequence version: 118 1118D58901CDF3FC cdk2_human Cyclin-dependent kinase 2 CDK2 p33 protein kinase Cell division protein kinase 2 CDKN2 protein protein human Homo sapiens Human MENFQKVEKIGEGTYGVVYKARNKLTGEVVALKKIRLDTETEGVPSTAIREISLLKELNHPNIVKLLDVIHTENKLYLVFEFLHQDLKKFMDASALTGIPLPLIKSYLFQLLQGLAFCHSHRVLHRDLKPQNLLINTEGAIKLADFGLARAFGVPVRTYTHEVVTLWYRAPEILLGCKYYSTAVDIWSLGCIFAEMVTRRALFPGDSEIDQLFRIFRTLGTPDEVVWPGVTSMPDYKPSFPKWARQDFSKVVPPLDEDGRSLLSQMLHYDPNKRISAKAALAHPFFQDVTKPVPHLRL DIP protein P24941 original sequence version: 142 DIP protein P24941 original sequence version: 118 DIP protein P24941 original sequence version: 143 DIP protein P24941 original sequence version: 144 DIP protein P24941 original sequence version: 152 DIP protein P24941 original sequence version: 182 DIP protein P24941-1 original sequence version: 204 DIP protein P24941 original sequence version: 90 DIP protein P24941 original sequence version: 190 DIP protein P24941 original sequence version: 168 F90A0F4E70910B51 ccne1_human G1/S-specific cyclin-E1 CCNE1 CCNE protein protein human Homo sapiens Human MPRERRERDAKERDTMKEDGGAEFSARSRKRKANVTVFLQDPDEEMAKIDRTARDQCGSQPWDNNAVCADPCSLIPTPDKEDDDRVYPNSTCKPRIIAPSRGSPLPVLSWANREEVWKIMLNKEKTYLRDQHFLEQHPLLQPKMRAILLDWLMEVCEVYKLHRETFYLAQDFFDRYMATQENVVKTLLQLIGISSLFIAAKLEEIYPPKLHQFAYVTDGACSGDEILTMELMIMKALKWRLSPLTIVSWLNVYMQVAYLNDLHEVLLPQYPQQIFIQIAELLDLCVLDVDCLEFPYGILAASALYHFSSSELMQKVSGYQWCDIENCVKWMVPFAMVIRETGSSKLKHFRGVADEDAHNIQTHRDSLDLLDKARAKKAMLSEQNRASPLPSGLLTPPQSGKKQSSGPEMA DIP protein P24864 original sequence version: 114 DIP protein P24864 original sequence version: 150 424DF0B253B7047E src-cdn1b-1 1 6 enzyme target enzyme target substrate substrate unspecified role unspecified role 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 P46527:p.Tyr74Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 14:56:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr74phe' Tue Jun 30 17:50:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr74Phe' P46527:p.Tyr88Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 18:34:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr88phe' Tue Jun 30 17:17:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr88Phe' P46527:p.Tyr89Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 15:30:10 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr89phe' Tue Jun 30 17:40:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr89Phe' ecoli Escherichia coli (strain K12) 5 enzyme enzyme unspecified role unspecified role spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection phosphorylation phosphorylation reaction phosphorylation PP1 1A 1B Src Preferentially Phosphorylates p27 at Y74 and Y88 In Vitro Src-mediated p27 phosphorylation was reduced by 36% and 33% in p27Y74F and p27Y88F, respectively, compared to p27 wild type, while that of p27Y89F was only reduced by 12% Phosphorylation of P46527 src-1 1 5 putative self putative self putative self putative self spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection phosphorylation phosphorylation reaction phosphorylation true Autophosphorylation of Src Src autophosphorylation indicates equal Src input (Figure 1B). 1B Phosphorylation of P12931 yes-cdn1b-1 1 6 enzyme target enzyme target substrate substrate unspecified role unspecified role P46527:p.Tyr74Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 15:46:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr74phe' Tue Jun 30 18:42:51 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr74Phe' P46527:p.Tyr88Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 20:19:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr88phe' Tue Jun 30 20:22:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr88Phe' P46527:p.Tyr89Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 18:25:54 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr89phe' Tue Jun 30 17:24:39 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr89Phe' 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 ecoli Escherichia coli (strain K12) 4 enzyme enzyme unspecified role unspecified role n-terminal 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection phosphorylation phosphorylation reaction phosphorylation 1D and 1E As for Src, Yes kinase assays showed that phosphorylation of p27 by Yes was reduced by mutational loss of Y74 and Y88 phosphorylation, while Y89F only modestly attenuated phosphorylation by Yes (Figures 1D and 1E). Yes-mediated p27 phosphorylation was substancially reduced in p27Y74F and p27Y88F, less so in p27Y89F phosphorylation of P46527 cdn1b-cdk2-1 3 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Similarly, Src transfection modestly reduced p27 in Cdk2 precipitates, while Cdk2- bound cyclin E was unaffected (Figure 3E). 2E cdn1b-cdk2-2 3 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role prey prey association association YFPp27WT immunoprecipitates contained less bound cyclin E and Cdk2 following Src transfection, and this was abrogated by PP1 (Figure 3D). 3D cdn1b-cdk2-3 2 7 unspecified role unspecified role prey prey spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 6 unspecified role unspecified role bait bait n-terminal 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 ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection association association Equal amounts of p27 from (B) were incubated with cyclin E-Cdk2. (C) p27, (D) Cdk2, and (E) cyclin E were precipitated and associated proteins blotted. 2C Phosphorylation of p27 (CDKN1B) by c-Src inhibits binding to CDK1 cdn1b-cdk2-4 2 7 unspecified role unspecified role bait bait spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 6 unspecified role unspecified role prey prey n-terminal 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 ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection association association Equal amounts of p27 from (B) were incubated with cyclin E-Cdk2. (C) p27, (D) Cdk2, and (E) cyclin E were precipitated and associated proteins blotted. 2D Phosphorylation of p27 (CDKN1B) by c-Src inhibits binding to CDK1 cdn1b-cdk2-5 2 6 unspecified role unspecified role prey prey n-terminal 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 ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 8 unspecified role unspecified role bait bait spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection association association Equal amounts of p27 from (B) were incubated with cyclin E-Cdk2. (C) p27, (D) Cdk2, and (E) cyclin E were precipitated and associated proteins blotted. 2E Phosphorylation of p27 (CDKN1B) by c-Src inhibits binding to CDK1