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/ Molecular basis for the specificity of p27 toward cyclin-dependent kinases that regulate cell division. Molecular basis for the specificity of p27 toward cyclin-dependent kinases that regulate cell division. J. Mol. Biol. (0022-2836) 2005 imex curation Lacy ER., Wang Y., Post J., Nourse A., Webb W., Mapelli M., Musacchio A., Siuzdak G., Kriwacki RW. Apoptosis - Interactions involving proteins with a function related to apoptosis p27 N15 labelled to enable peptide identification. Proteins incubated with a mixture of trypsin and chymotrysin and reaction products were analyzed using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Identity of Cyclin A2 taken from sequence length. Cyclin A is a known synonym of Cyclin A2. SOurce of CDK5 and p25 taken from PMID: 11583627 richard.kriwacki@stjude.org 26-APR-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Lacy ER., Wang Y., Post J., Nourse A., Webb W., Mapelli M., Musacchio A., Siuzdak G., Kriwacki RW. Apoptosis - Interactions involving proteins with a function related to apoptosis p27 N15 labelled to enable peptide identification. Proteins incubated with a mixture of trypsin and chymotrysin and reaction products were analyzed using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Identity of Cyclin A2 taken from sequence length. Cyclin A is a known synonym of Cyclin A2. SOurce of CDK5 and p25 taken from PMID: 11583627 richard.kriwacki@stjude.org Accepted 2007-MAY-12 by DAVET. 26-APR-2007: Contacted by JYOTI. J. Mol. Biol. (0022-2836) 2005 Only protein-protein interactions imex curation Molecular basis for the specificity of p27 toward cyclin-dependent kinases that regulate cell division. Molecular basis for the specificity of p27 toward cyclin-dependent kinases that regulate cell division. J. Mol. Biol. (0022-2836) 2005 imex curation Lacy ER., Wang Y., Post J., Nourse A., Webb W., Mapelli M., Musacchio A., Siuzdak G., Kriwacki RW. Apoptosis - Interactions involving proteins with a function related to apoptosis p27 N15 labelled to enable peptide identification. Proteins incubated with a mixture of trypsin and chymotrysin and reaction products were analyzed using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Identity of Cyclin A2 taken from sequence length. Cyclin A is a known synonym of Cyclin A2. SOurce of CDK5 and p25 taken from PMID: 11583627 richard.kriwacki@stjude.org 26-APR-2007: Contacted by JYOTI. 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 Lacy ER., Wang Y., Post J., Nourse A., Webb W., Mapelli M., Musacchio A., Siuzdak G., Kriwacki RW. Apoptosis - Interactions involving proteins with a function related to apoptosis p27 N15 labelled to enable peptide identification. Proteins incubated with a mixture of trypsin and chymotrysin and reaction products were analyzed using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Identity of Cyclin A2 taken from sequence length. Cyclin A is a known synonym of Cyclin A2. SOurce of CDK5 and p25 taken from PMID: 11583627 richard.kriwacki@stjude.org Accepted 2007-MAY-12 by DAVET. 26-APR-2007: Contacted by JYOTI. J. Mol. Biol. (0022-2836) 2005 Only protein-protein interactions imex curation Molecular basis for the specificity of p27 toward cyclin-dependent kinases that regulate cell division. Molecular basis for the specificity of p27 toward cyclin-dependent kinases that regulate cell division. J. Mol. Biol. (0022-2836) 2005 imex curation Lacy ER., Wang Y., Post J., Nourse A., Webb W., Mapelli M., Musacchio A., Siuzdak G., Kriwacki RW. Apoptosis - Interactions involving proteins with a function related to apoptosis p27 N15 labelled to enable peptide identification. Proteins incubated with a mixture of trypsin and chymotrysin and reaction products were analyzed using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Identity of Cyclin A2 taken from sequence length. Cyclin A is a known synonym of Cyclin A2. SOurce of CDK5 and p25 taken from PMID: 11583627 richard.kriwacki@stjude.org 26-APR-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro footprinting footprinting fingerprinting peptide massfingerprinting fingerprinting Lacy ER., Wang Y., Post J., Nourse A., Webb W., Mapelli M., Musacchio A., Siuzdak G., Kriwacki RW. Apoptosis - Interactions involving proteins with a function related to apoptosis p27 N15 labelled to enable peptide identification. Proteins incubated with a mixture of trypsin and chymotrysin and reaction products were analyzed using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Identity of Cyclin A2 taken from sequence length. Cyclin A is a known synonym of Cyclin A2. SOurce of CDK5 and p25 taken from PMID: 11583627 richard.kriwacki@stjude.org Accepted 2007-MAY-12 by DAVET. 26-APR-2007: Contacted by JYOTI. J. Mol. Biol. (0022-2836) 2005 Only protein-protein interactions imex curation 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 q15078-pro_0000004795 Cyclin-dependent kinase 5 activator 1, p25 CDK5R1 CDK5R NCK5A Cyclin-dependent kinase 5 regulatory subunit 1 TPKII regulatory subunit protein protein human Homo sapiens Human AQPPPAQPPAPPASQLSGSQTGGSSSVKKAPHPAVTSAGTPKRVIVQASTSELLRCLGEFLCRRCYRLKHLSPTDPVLWLRSVDRSLLLQGWQDQGFITPANVVFLYMLCRDVISSEVGSDHELQAVLLTCLYLSYSYMGNEISYPLKPFLVESCKEAFWDRCLSVINLMSSKMLQINADPHYFTQVFSDLKNESGQEDKKRLLLGLDR 99 307 DC5A740D6E9F6DBF ccna2_human Cyclin-A2 Cyclin A CCNA2 CCN1 CCNA protein protein human Homo sapiens Human MLGNSAPGPATREAGSALLALQQTALQEDQENINPEKAAPVQQPRTRAALAVLKSGNPRGLAQQQRPKTRRVAPLKDLPVNDEHVTVPPWKANSKQPAFTIHVDEAEKEAQKKPAESQKIEREDALAFNSAISLPGPRKPLVPLDYPMDGSFESPHTMDMSIILEDEKPVSVNEVPDYHEDIHTYLREMEVKCKPKVGYMKKQPDITNSMRAILVDWLVEVGEEYKLQNETLHLAVNYIDRFLSSMSVLRGKLQLVGTAAMLLASKFEEIYPPEVAEFVYITDDTYTKKQVLRMEHLVLKVLTFDLAAPTVNQFLTQYFLHQQPANCKVESLAMFLGELSLIDADPYLKYLPSVIAGAAFHLALYTVTGQSWPESLIRKTGYTLESLKPCLMDLHQTYLKAPQHAQQSIREKYKNSKYHGVSLLNPPETLNL DIP protein P20248 original sequence version: 183 DIP protein P20248 original sequence version: 146 DIP protein P20248 original sequence version: 168 DIP protein P20248 original sequence version: 149 DIP protein P20248 original sequence version: 111 97763A2897DD35F4 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 cdk5_human Cyclin-dependent-like kinase 5 CDK5 Cell division protein kinase 5 Tau protein kinase II catalytic subunit Serine/threonine-protein kinase PSSALRE CDKN5 protein protein human Homo sapiens Human MQKYEKLEKIGEGTYGTVFKAKNRETHEIVALKRVRLDDDDEGVPSSALREICLLKELKHKNIVRLHDVLHSDKKLTLVFEFCDQDLKKYFDSCNGDLDPEIVKSFLFQLLKGLGFCHSRNVLHRDLKPQNLLINRNGELKLADFGLARAFGIPVRCYSAEVVTLWYRPPDVLFGAKLYSTSIDMWSAGCIFAELANAGRPLFPGNDVDDQLKRIFRLLGTPTEEQWPSMTKLPDYKPYPMYPATTSLVNVVPKLNATGRDLLQNLLKCNPVQRISAEEALQHPYFSDFCPP DIP protein Q00535 original sequence version: 129 DIP protein Q00535 original sequence version: 140 54D10495F017D527 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 cdkn1b-ccna2-1 2 7 unspecified role unspecified role bait bait 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 kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 6 unspecified role unspecified role prey prey 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 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 13 16 physical association physical association Association (ka) and dissociation (kd) rate constants for the binding of CKIs to components of Cdk/cyclin complexes derived from the analysis of SPR data at 25C Table 1 ka (slow) 21.40 +/- 2.6 exp-4 M-1 s-1, ka (fast) 1.12 +/- 0.11 exp6 M-1 s-1, kd (slow) 4.70 +/- 0.11 exp-4 s-1, kd (fast)4.10 +/- 0.31 exp-2 s-1 at 25C. The binding of Cdk2/cyclin A to p27-KID exhibited biphasic kinetics. cdkn1b-ccna2-2 2 6 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) 7 unspecified role unspecified role bait bait kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) 4 unspecified role unspecified role prey prey 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 ecobb Escherichia coli (strain B / BL21) physical association physical association Association (ka) and dissociation (kd) rate constants for the binding of CKIs to components of Cdk/cyclin complexes derived from the analysis of SPR data at 25C Table 1 ka (slow) 0.56 +/- 0.11 exp-4 M-1 s-1, ka (fast) 1.57 +/- 0.04 exp6 M-1 s-1, kd (slow) 0.02 +/- 0.001 exp-4 s-1, kd (fast)1.38 +/- 0.07 exp-2 s-1 at 25C. The binding of Cdk2/cyclin A to p27-KID exhibited biphasic kinetics. cdkn1b-cdk2-6 2 7 unspecified role unspecified role bait bait kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) 4 unspecified role unspecified role prey prey 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Association (ka) and dissociation (kd) rate constants for the binding of CKIs to components of Cdk/cyclin complexes derived from the analysis of SPR data at 25C Table 1 ka (slow) 0.26 +/- 0.02 exp-4 M-1 s-1, kd (slow) 3.82 +/- 0.93 exp-4 s-1 at 25C. The binding of Cdk2 to p27-KID exhibited single phase kinetics. cdkn1b-cdk5-1 2 8 unspecified role unspecified role prey prey 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 Q00535:p.Asp144Asn mutation mutation certain certain sequence position certain certain certain sequence position certain D N Wed May 02 15:32:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp144asn' Tue Jun 30 16:05:48 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp144Asn' ecobb Escherichia coli (strain B / BL21) 7 unspecified role unspecified role bait bait kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Association (ka) and dissociation (kd) rate constants for the binding of CKIs to components of Cdk/cyclin complexes derived from the analysis of SPR data at 25C Table 1 ka (slow) 0.45 +/- 0.08 exp-4 M-1 s-1, kd (slow) 2.07 +/- 0.1 exp-4 s-1 at 25C. The binding of Cdk5 to p27-KID exhibited single phase kinetics. CDK5 is mutated to a kinase-dead version cdk5r1-cdk5-1 2 5 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 prey prey 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 Q00535:p.Asp144Asn mutation mutation certain certain sequence position certain certain certain sequence position certain D N Wed May 02 15:19:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp144asn' Tue Jun 30 16:30:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp144Asn' 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 7 unspecified role unspecified role bait bait kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) physical association physical association Association (ka) and dissociation (kd) rate constants for the binding of CKIs to components of Cdk/cyclin complexes derived from the analysis of SPR data at 25C Table 1 ka (slow) 0.37 +/- 0.05 exp-4 M-1 s-1, kd (slow) 1.83 +/- 0.72 exp-4 s-1 at 25C. The binding of Cdk5 to p27-KID exhibited single phase kinetics. CDK5 is mutated to a kinase-dead version cdkn1a-ccna1-2 2 9 unspecified role unspecified role bait bait 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 kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 6 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) physical association physical association Association (ka) and dissociation (kd) rate constants for the binding of CKIs to components of Cdk/cyclin complexes derived from the analysis of SPR data at 25C Table 1 ka (slow) 3.85 +/- 0.24 exp-4 M-1 s-1, ka (fast) 2.04 +/- 1.25 exp6 M-1 s-1, kd (slow) 7.03 +/- 1.92 exp-4 s-1, kd (fast)2.69 +/- 0.29 exp-2 s-1 at 25C. The binding of Cdk2/cyclin A to p21-KID exhibited biphasic kinetics. cdkn1a-ccna2-1 2 9 unspecified role unspecified role bait bait 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 kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 4 unspecified role unspecified role prey prey 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 ecobb Escherichia coli (strain B / BL21) 6 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) physical association physical association Association (ka) and dissociation (kd) rate constants for the binding of CKIs to components of Cdk/cyclin complexes derived from the analysis of SPR data at 25C Table 1 ka (slow) 3.95 +/- 1.06 exp-4 M-1 s-1, ka (fast) 0.56 +/- 0.1 exp6 M-1 s-1, kd (slow) 0.98 +/- 0.74 exp-4 s-1, kd (fast) 0.03 +/- 0.003 exp-2 s-1 at 25C. The binding of Cdk2/cyclin A to p21-KID exhibited biphasic kinetics. cdkn1a-cdk2-3 2 9 unspecified role unspecified role bait bait 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 kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 4 unspecified role unspecified role prey prey 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Association (ka) and dissociation (kd) rate constants for the binding of CKIs to components of Cdk/cyclin complexes derived from the analysis of SPR data at 25C Table 1 ka (slow) 1.06 +/- 0.13 exp-4 M-1 s-1, kd (slow) 1.43 +/- 0.21 exp-4 s-1 at 25C. The binding of Cdk2 to p21-KID exhibited single phase kinetics. cdkn1a-cdk5-1 2 9 unspecified role unspecified role bait bait kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) 8 unspecified role unspecified role prey prey Q00535:p.Asp144Asn mutation mutation certain certain sequence position certain certain certain sequence position certain D N Wed May 02 18:06:45 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp144asn' Tue Jun 30 19:20:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp144Asn' 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Association (ka) and dissociation (kd) rate constants for the binding of CKIs to components of Cdk/cyclin complexes derived from the analysis of SPR data at 25C Table 1 ka (slow) 2.12 +/- 0.51 exp-4 M-1 s-1, kd (slow) 3.19 +/- 0.77 exp-4 s-1 at 25C. The binding of Cdk5 to p21-KID exhibited single phase kinetics. CDK5 is mutated to a kinase-dead version cdk5r1-cdk5-2 2 9 unspecified role unspecified role bait bait 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 kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 8 unspecified role unspecified role prey prey Q00535:p.Asp144Asn mutation mutation certain certain sequence position certain certain certain sequence position certain D N Wed May 02 15:36:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp144asn' Tue Jun 30 16:52:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp144Asn' 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 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 5 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 physical association physical association Association (ka) and dissociation (kd) rate constants for the binding of CKIs to components of Cdk/cyclin complexes derived from the analysis of SPR data at 25C Table 1 ka (slow) 1.52 +/- 0.48 exp-4 M-1 s-1, kd (slow) 6.74 +/- 1.46 exp-4 s-1 at 25C. The binding of Cdk5 to p21-KID exhibited single phase kinetics. CDK5 is mutated to a kinase-dead version cdkn1b-ccna2-3 3 6 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) 7 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 kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) physical association physical association Trypsin cleavage patterns based on analysis by MALDI-TOF mass spectrometry for (a) free p27-KID, and (b) p27-KID in the presence of cyclin A, (c) Cdk2, (d) Cdk5, (e) Cdk5/p25 and (f) Cdk2/cyclin A. Figure 3 Arg58 and Lys59 were cleaved when Cyclin A2 (CCNA2) was bound to p27 (CDKN1B). cdkn1b-ccna2-4 3 4 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 ecobb Escherichia coli (strain B / BL21) 7 unspecified role unspecified role neutral component neutral component kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) 6 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) physical association physical association Trypsin cleavage patterns based on analysis by MALDI-TOF mass spectrometry for (a) free p27-KID, and (b) p27-KID in the presence of cyclin A, (c) Cdk2, (d) Cdk5, (e) Cdk5/p25 and (f) Cdk2/cyclin A. p27 is largely protected from cleavage when bound to CCNA2/CDK2 Figure 3 cdkn1b-cdk2-7 3 4 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 ecobb Escherichia coli (strain B / BL21) 7 unspecified role unspecified role neutral component neutral component region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Trypsin cleavage patterns based on analysis by MALDI-TOF mass spectrometry for (a) free p27-KID, and (b) p27-KID in the presence of cyclin A, (c) Cdk2, (d) Cdk5, (e) Cdk5/p25 and (f) Cdk2/cyclin A. Figure 3 P27 is largely protected from cleavage when bound to CDK2 cdkn1b-cdk5-2 3 7 unspecified role unspecified role neutral component neutral component region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) 8 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 Q00535:p.Asp144Asn mutation mutation certain certain sequence position certain certain certain sequence position certain D N Wed May 02 17:40:40 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp144asn' Tue Jun 30 18:14:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp144Asn' ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Trypsin cleavage patterns based on analysis by MALDI-TOF mass spectrometry for (a) free p27-KID, and (b) p27-KID in the presence of cyclin A, (c) Cdk2, (d) Cdk5, (e) Cdk5/p25 and (f) Cdk2/cyclin A. Figure 3 p27 is largely protected from cleavage when bound to CDK5 CDK5 is mutated to a kinase-dead version cdk5r1-cdk5-3 3 7 unspecified role unspecified role neutral component neutral component region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) 5 unspecified role unspecified role neutral component neutral component 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 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 Q00535:p.Asp144Asn mutation mutation certain certain sequence position certain certain certain sequence position certain D N Wed May 02 19:20:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp144asn' Tue Jun 30 15:58:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp144Asn' 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 physical association physical association Trypsin cleavage patterns based on analysis by MALDI-TOF mass spectrometry for (a) free p27-KID, and (b) p27-KID in the presence of cyclin A, (c) Cdk2, (d) Cdk5, (e) Cdk5/p25 and (f) Cdk2/cyclin A. Figure 3 CDK5 is mutated to a kinase-dead version p27-KID is largely protected from cleavage when bound to p25/CDK5 cdkn1b-ccna2-5 1 6 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) 7 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 kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 128 132 direct interaction direct interaction NMR data further support the results of SPR and limited proteolysis experiments, by showing that a 13 amino acid residue peptide corresponding to domain 1 of p27 bound to cyclin A and Cdk2/cylin A but not to Cdk5/p25. Figure 4A cdkn1b-ccna2-6 1 6 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) 7 unspecified role unspecified role neutral component neutral component kid region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobb Escherichia coli (strain B / BL21) 4 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 ecobb Escherichia coli (strain B / BL21) physical association physical association NMR data further support the results of SPR and limited proteolysis experiments, by showing that a 13 amino acid residue peptide corresponding to domain 1 of p27 bound to cyclin A and Cdk2/cylin A but not to Cdk5/p25. Figure 4B