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/ Structural and biochemical studies of human proliferating cell nuclear antigen complexes provide a rationale for cyclin association and inhibitor design. Structural and biochemical studies of human proliferating cell nuclear antigen complexes provide a rationale for cyclin association and inhibitor design. Proceedings of the National Academy of Sciences of the United States of America 2005 imex curation Kontopidis G., Wu SY., Zheleva DI., Taylor P., McInnes C., Lane DP., Fischer PM., Walkinshaw MD. m.walkinshaw@ed.ac.uk Only protein-protein interactions imex curation in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Kontopidis G., Wu SY., Zheleva DI., Taylor P., McInnes C., Lane DP., Fischer PM., Walkinshaw MD. m.walkinshaw@ed.ac.uk Accepted 2021-AUG-31 AT 15:33 BST AT 15:33 BST by MGT Proceedings of the National Academy of Sciences of the United States of America 2005 Only protein-protein interactions imex curation imex curation Structural and biochemical studies of human proliferating cell nuclear antigen complexes provide a rationale for cyclin association and inhibitor design. Structural and biochemical studies of human proliferating cell nuclear antigen complexes provide a rationale for cyclin association and inhibitor design. Proceedings of the National Academy of Sciences of the United States of America 2005 imex curation Kontopidis G., Wu SY., Zheleva DI., Taylor P., McInnes C., Lane DP., Fischer PM., Walkinshaw MD. m.walkinshaw@ed.ac.uk Only protein-protein interactions imex curation in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Kontopidis G., Wu SY., Zheleva DI., Taylor P., McInnes C., Lane DP., Fischer PM., Walkinshaw MD. For the Rb protein the species origin has been assumed to be Human based on sequence length. m.walkinshaw@ed.ac.uk Accepted 2021-AUG-31 AT 15:33 BST AT 15:33 BST by MGT Proceedings of the National Academy of Sciences of the United States of America 2005 Only protein-protein interactions imex curation imex curation inhibitor peptide inhibitor peptide SAVLQKKITDYFHPKK peptide peptide polypeptide oligopeptide oligopeptide polypeptide synthetic construct synthetic construct SAVLQKKITDYFHPKK ccnd1_human G1/S-specific cyclin-D1 CCND1 BCL1 PRAD1 PRAD1 oncogene BCL-1 oncogene B-cell lymphoma 1 protein protein protein human Homo sapiens Human MEHQLLCCEVETIRRAYPDANLLNDRVLRAMLKAEETCAPSVSYFKCVQKEVLPSMRKIVATWMLEVCEEQKCEEEVFPLAMNYLDRFLSLEPVKKSRLQLLGATCMFVASKMKETIPLTAEKLCIYTDNSIRPEELLQMELLLVNKLKWNLAAMTPHDFIEHFLSKMPEAEENKQIIRKHAQTFVALCATDVKFISNPPSMVAAGSVVAAVQGLNLRSPNNFLSYYRLTRFLSRVIKCDPDCLRACQEQIEALLESSLRQAQQNMDPKAAEEEEEEEEEVDLACTPTDVRDVDI DIP protein P24385 original sequence version: 163 DIP protein P24385 original sequence version: 165 DIP protein P24385 original sequence version: 160 DIP protein P24385 original sequence version: 121 DIP protein P24385 original sequence version: 122 DIP protein P24385 original sequence version: 143 DIP protein P24385 original sequence version: 127 3CC00C9905F58D3A cdk4_human Cyclin-dependent kinase 4 CDK4 Cell division protein kinase 4 PSK-J3 protein protein human Homo sapiens Human MATSRYEPVAEIGVGAYGTVYKARDPHSGHFVALKSVRVPNGGGGGGGLPISTVREVALLRRLEAFEHPNVVRLMDVCATSRTDREIKVTLVFEHVDQDLRTYLDKAPPPGLPAETIKDLMRQFLRGLDFLHANCIVHRDLKPENILVTSGGTVKLADFGLARIYSYQMALTPVVVTLWYRAPEVLLQSTYATPVDMWSVGCIFAEMFRRKPLFCGNSEADQLGKIFDLIGLPPEDDWPRDVSLPRGAFPPRGPRPVQSVVPEMEESGAQLLLEMLTFNPHKRISAFRALQHSYLHKDEGNPE DIP protein P11802 original sequence version: 177 DIP protein P11802 original sequence version: 135 DIP protein P11802 original sequence version: 136 DIP protein P11802 original sequence version: 200 DIP protein P11802 original sequence version: 86 0916A0C07403A33A rb_human Retinoblastoma-associated protein pRb pp110 p105-Rb RB1 p110-RB1 protein protein human Homo sapiens Human MPPKTPRKTAATAAAAAAEPPAPPPPPPPEEDPEQDSGPEDLPLVRLEFEETEEPDFTALCQKLKIPDHVRERAWLTWEKVSSVDGVLGGYIQKKKELWGICIFIAAVDLDEMSFTFTELQKNIEISVHKFFNLLKEIDTSTKVDNAMSRLLKKYDVLFALFSKLERTCELIYLTQPSSSISTEINSALVLKVSWITFLLAKGEVLQMEDDLVISFQLMLCVLDYFIKLSPPMLLKEPYKTAVIPINGSPRTPRRGQNRSARIAKQLENDTRIIEVLCKEHECNIDEVKNVYFKNFIPFMNSLGLVTSNGLPEVENLSKRYEEIYLKNKDLDARLFLDHDKTLQTDSIDSFETQRTPRKSNLDEEVNVIPPHTPVRTVMNTIQQLMMILNSASDQPSENLISYFNNCTVNPKESILKRVKDIGYIFKEKFAKAVGQGCVEIGSQRYKLGVRLYYRVMESMLKSEEERLSIQNFSKLLNDNIFHMSLLACALEVVMATYSRSTSQNLDSGTDLSFPWILNVLNLKAFDFYKVIESFIKAEGNLTREMIKHLERCEHRIMESLAWLSDSPLFDLIKQSKDREGPTDHLESACPLNLPLQNNHTAADMYLSPVRSPKKKGSTTRVNSTANAETQATSAFQTQKPLKSTSLSLFYKKVYRLAYLRLNTLCERLLSEHPELEHIIWTLFQHTLQNEYELMRDRHLDQIMMCSMYGICKVKNIDLKFKIIVTAYKDLPHAVQETFKRVLIKEEEYDSIIVFYNSVFMQRLKTNILQYASTRPPTLSPIPHIPRSPYKFPSSPLRIPGGNIYISPLKSPYKISEGLPTPTKMTPRSRILVSIGESFGTSEKFQKINQMVCNSDRVLKRSAEGSNPPKPLKKLRFDIEGSDEADGSKHLPGESKFQQKLAEMTSTRTRMQKQKMNDSMDTSNKEEK DIP protein P06400 original sequence version: 102 DIP protein P06400 original sequence version: 195 DIP protein P06400 original sequence version: 204 DIP protein P06400 original sequence version: 126 DIP protein P06400 original sequence version: 151 DIP protein P06400 original sequence version: 153 DIP protein P06400 original sequence version: 181 DIP protein P06400 original sequence version: 184 DIP protein P06400 original sequence version: 213 DIP protein P06400 original sequence version: 208 DIP protein P06400 original sequence version: 174 DIP protein P06400 original sequence version: 212 DIP protein P06400 original sequence version: 179 DIP protein P06400 original sequence version: 146 C8E746111E19CC32 pcna_human Proliferating cell nuclear antigen PCNA Cyclin protein protein human Homo sapiens Human MFEARLVQGSILKKVLEALKDLINEACWDISSSGVNLQSMDSSHVSLVQLTLRSEGFDTYRCDRNLAMGVNLTSMSKILKCAGNEDIITLRAEDNADTLALVFEAPNQEKVSDYEMKLMDLDVEQLGIPEQEYSCVVKMPSGEFARICRDLSHIGDAVVISCAKDGVKFSASGELGNGNIKLSQTSNVDKEEEAVTIEMNEPVQLTFALRYLNFFTKATPLSSTVTLSMSADVPLVVEYKIADMGHLKYYLAPKIEDEEGS DIP protein P12004 original sequence version: 200 DIP protein P12004 original sequence version: 128 DIP protein P12004 original sequence version: 139 DIP protein P12004 original sequence version: 185 DIP protein P12004 original sequence version: 102 DIP protein P12004 original sequence version: 105 DIP protein P12004 original sequence version: 158 E6F08E7EDBC48B00 cdn1a_human Cyclin-dependent kinase inhibitor 1 CDKN1A CAP20 CDKN1 CIP1 MDA6 PIC1 SDI1 WAF1 p21 CDK-interacting protein 1 Melanoma differentiation-associated protein 6 protein protein human Homo sapiens Human MSEPAGDVRQNPCGSKACRRLFGPVDSEQLSRDCDALMAGCIQEARERWNFDFVTETPLEGDFAWERVRGLGLPKLYLPTGPRRGRDELGGGRRPGTSPALLQGTAEEDHVDLSLSCTLVPRSGEQAEGSPGGPGDSQGRKRRQTSMTDFYHSKRRLIFSKRKP DIP protein P38936 original sequence version: 120 DIP protein P38936 original sequence version: 165 DIP protein P38936 original sequence version: 160 98D1E7C519ADFCA9 pcna-13 1 7 unspecified role unspecified role neutral component neutral component purified purified ecobd Escherichia coli (strain B / BL21-DE3) direct interaction direct interaction Table 1 pcna-14 1 7 unspecified role unspecified role neutral component neutral component purified purified ecobd Escherichia coli (strain B / BL21-DE3) 3 unspecified role unspecified role neutral component neutral component purified purified inhibitor peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Fig.2, Table 1 ccnd1-cdk4-27 2 4 enzyme regulator enzyme regulator neutral component neutral component purified purified cyclin-D1 G1/S-specific cyclin-D1 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobd Escherichia coli (strain B / BL21-DE3) 5 enzyme enzyme neutral component neutral component purified purified tag 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 CDK4 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 enzyme target enzyme target substrate substrate neutral component neutral component purified purified hyperphosphorylation domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain GST-tag 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 region phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined resulting-ptm resulting-ptm resulting ptm resulting-ptm ecobd Escherichia coli (strain B / BL21-DE3) For the Rb protein the species origin has been assumed to be Human based on sequence length. 17 20 phosphorylation phosphorylation reaction phosphorylation Fig. 5 ccnd1-cdk4-28 2 8 inhibitor inhibitor neutral component neutral component purified purified Inhibitory region of cyclin-dependent kinase inhibitor 1 Inhibitory region of cyclin-dependent kinase inhibitor 1 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 4 enzyme regulator enzyme regulator neutral component neutral component purified purified cyclin-D1 G1/S-specific cyclin-D sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobd Escherichia coli (strain B / BL21-DE3) 5 enzyme enzyme neutral component neutral component purified purified tag 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 CDK4 Cyclin-dependent kinase 4 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 enzyme target enzyme target substrate substrate neutral component neutral component purified purified hyperphosphorylation domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain GST-tag 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 region phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined resulting-ptm resulting-ptm resulting ptm resulting-ptm ecobd Escherichia coli (strain B / BL21-DE3) For the Rb protein the species origin has been assumed to be Human based on sequence length. 32 29 phosphorylation phosphorylation reaction phosphorylation Fig. 5 ccnd1-cdk4-29 2 7 inhibitor inhibitor neutral component neutral component purified purified PCNA Proliferating cell nuclear antigen sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 8 inhibitor inhibitor neutral component neutral component purified purified Inhibitory region of cyclin-dependent kinase inhibitor 1 Inhibitory region of cyclin-dependent kinase inhibitor 1 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 4 enzyme regulator enzyme regulator neutral component neutral component purified purified cyclin-D1 G1/S-specific cyclin-D1 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobd Escherichia coli (strain B / BL21-DE3) 5 enzyme enzyme neutral component neutral component purified purified CDK4 Cyclin-dependent kinase 4 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain tag 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 6 enzyme target enzyme target substrate substrate neutral component neutral component purified purified hyperphosphorylation domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain tag 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 region phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined resulting-ptm resulting-ptm resulting ptm resulting-ptm ecobd Escherichia coli (strain B / BL21-DE3) For the Rb protein the species origin has been assumed to be Human based on sequence length. 43 45 phosphorylation phosphorylation reaction phosphorylation Fig. 5b-d Concentrations of PCNA > 15uM resulted in decreased pRb phosphorylation.