mbinfo MBInfo http://www.mechanobio.info/ Direct association of pp125FAK with paxillin, the focal adhesion-targeting mechanism of pp125FAK. Direct association of pp125FAK with paxillin, the focal adhesion-targeting mechanism of pp125FAK. J. Exp. Med. (0022-1007) 1995 imex curation Tachibana K., Sato T., D'Avirro N., Morimoto C. Inferred from the paper, the GST-FAK fusion proteins were produced from E.coli JM105 cells; and the paxillin proteins were from either a human T lymphoblastoid cell line, HPB-ALL, or a human breast cancer cell line, T-47D. The contact email is not given by the author in the paper. 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting autoradiography autoradiography Tachibana K., Sato T., D'Avirro N., Morimoto C. Inferred from the paper, the GST-FAK fusion proteins were produced from E.coli JM105 cells; and the paxillin proteins were from either a human T lymphoblastoid cell line, HPB-ALL, or a human breast cancer cell line, T-47D. The contact email is not given by the author in the paper. Accepted 2012-JUL-24 by ORCHARD 19-Sept-2012: Contacted by IntAct-Help. J. Exp. Med. (0022-1007) 1995 Only protein-protein interactions imex curation Direct association of pp125FAK with paxillin, the focal adhesion-targeting mechanism of pp125FAK. Direct association of pp125FAK with paxillin, the focal adhesion-targeting mechanism of pp125FAK. J. Exp. Med. (0022-1007) 1995 imex curation Tachibana K., Sato T., D'Avirro N., Morimoto C. Inferred from the paper, the GST-FAK fusion proteins were produced from E.coli JM105 cells; and the paxillin proteins were from either a human T lymphoblastoid cell line, HPB-ALL, or a human breast cancer cell line, T-47D. The contact email is not given by the author in the paper. 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Tachibana K., Sato T., D'Avirro N., Morimoto C. Inferred from the paper, the GST-FAK fusion proteins were produced from E.coli JM105 cells; and the paxillin proteins were from either a human T lymphoblastoid cell line, HPB-ALL, or a human breast cancer cell line, T-47D. The contact email is not given by the author in the paper. Accepted 2012-JUL-24 by ORCHARD 19-Sept-2012: Contacted by IntAct-Help. J. Exp. Med. (0022-1007) 1995 Only protein-protein interactions imex curation Direct association of pp125FAK with paxillin, the focal adhesion-targeting mechanism of pp125FAK. Direct association of pp125FAK with paxillin, the focal adhesion-targeting mechanism of pp125FAK. J. Exp. Med. (0022-1007) 1995 imex curation Tachibana K., Sato T., D'Avirro N., Morimoto C. Inferred from the paper, the GST-FAK fusion proteins were produced from E.coli JM105 cells; and the paxillin proteins were from either a human T lymphoblastoid cell line, HPB-ALL, or a human breast cancer cell line, T-47D. The contact email is not given by the author in the paper. 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions human-hpb-all Homo sapiens hpb-all Human T lymphoblastoid cell line HPB-ALL This cell line has been shown to have been contaminated by Jurkat cells at some unknown point in its usage. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Tachibana K., Sato T., D'Avirro N., Morimoto C. Inferred from the paper, the GST-FAK fusion proteins were produced from E.coli JM105 cells; and the paxillin proteins were from either a human T lymphoblastoid cell line, HPB-ALL, or a human breast cancer cell line, T-47D. The contact email is not given by the author in the paper. Accepted 2012-JUL-24 by ORCHARD 19-Sept-2012: Contacted by IntAct-Help. J. Exp. Med. (0022-1007) 1995 Only protein-protein interactions imex curation Direct association of pp125FAK with paxillin, the focal adhesion-targeting mechanism of pp125FAK. Direct association of pp125FAK with paxillin, the focal adhesion-targeting mechanism of pp125FAK. J. Exp. Med. (0022-1007) 1995 imex curation Tachibana K., Sato T., D'Avirro N., Morimoto C. Inferred from the paper, the GST-FAK fusion proteins were produced from E.coli JM105 cells; and the paxillin proteins were from either a human T lymphoblastoid cell line, HPB-ALL, or a human breast cancer cell line, T-47D. The contact email is not given by the author in the paper. 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions human-t47d human breast cancer tumour cell line-epithelial cells t47d Epithelial cells derived from human breast tumour anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Tachibana K., Sato T., D'Avirro N., Morimoto C. Inferred from the paper, the GST-FAK fusion proteins were produced from E.coli JM105 cells; and the paxillin proteins were from either a human T lymphoblastoid cell line, HPB-ALL, or a human breast cancer cell line, T-47D. The contact email is not given by the author in the paper. Accepted 2012-JUL-24 by ORCHARD 19-Sept-2012: Contacted by IntAct-Help. J. Exp. Med. (0022-1007) 1995 Only protein-protein interactions imex curation fak1_human Focal adhesion kinase 1 PTK2 FAK FAK1 pp125FAK Protein-tyrosine kinase 2 Focal adhesion kinase-related nonkinase Protein phosphatase 1 regulatory subunit 71 p125FAK protein protein human Homo sapiens Human MAAAYLDPNLNHTPNSSTKTHLGTGMERSPGAMERVLKVFHYFESNSEPTTWASIIRHGDATDVRGIIQKIVDSHKVKHVACYGFRLSHLRSEEVHWLHVDMGVSSVREKYELAHPPEEWKYELRIRYLPKGFLNQFTEDKPTLNFFYQQVKSDYMLEIADQVDQEIALKLGCLEIRRSYWEMRGNALEKKSNYEVLEKDVGLKRFFPKSLLDSVKAKTLRKLIQQTFRQFANLNREESILKFFEILSPVYRFDKECFKCALGSSWIISVELAIGPEEGISYLTDKGCNPTHLADFTQVQTIQYSNSEDKDRKGMLQLKIAGAPEPLTVTAPSLTIAENMADLIDGYCRLVNGTSQSFIIRPQKEGERALPSIPKLANSEKQGMRTHAVSVSETDDYAEIIDEEDTYTMPSTRDYEIQRERIELGRCIGEGQFGDVHQGIYMSPENPALAVAIKTCKNCTSDSVREKFLQEALTMRQFDHPHIVKLIGVITENPVWIIMELCTLGELRSFLQVRKYSLDLASLILYAYQLSTALAYLESKRFVHRDIAARNVLVSSNDCVKLGDFGLSRYMEDSTYYKASKGKLPIKWMAPESINFRRFTSASDVWMFGVCMWEILMHGVKPFQGVKNNDVIGRIENGERLPMPPNCPPTLYSLMTKCWAYDPSRRPRFTELKAQLSTILEEEKAQQEERMRMESRRQATVSWDSGGSDEAPPKPSRPGYPSPRSSEGFYPSPQHMVQTNHYQVSGYPGSHGITAMAGSIYPGQASLLDQTDSWNHRPQEIAMWQPNVEDSTVLDLRGIGQVLPTHLMEERLIRQQQEMEEDQRWLEKEERFLKPDVRLSRGSIDREDGSLQGPIGNQHIYQPVGKPDPAAPPKKPPRPGAPGHLGSLASLSSPADSYNEGVKLQPQEISPPPTANLDRSNDKVYENVTGLVKAVIEMSSKIQPAPPEEYVPMVKEVGLALRTLLATVDETIPLLPASTHREIEMAQKLLNSDLGELINKMKLAQQYVMTSLQQEYKKQMLTAAHALAVDAKNLLDVIDQARLKMLGQTRPH D8A4C15138AB0243 crk_human Adapter molecule crk Proto-oncogene c-Crk CRK p38 protein protein human Homo sapiens Human MAGNFDSEERSSWYWGRLSRQEAVALLQGQRHGVFLVRDSSTSPGDYVLSVSENSRVSHYIINSSGPRPPVPPSPAQPPPGVSPSRLRIGDQEFDSLPALLEFYKIHYLDTTTLIEPVSRSRQGSGVILRQEEAEYVRALFDFNGNDEEDLPFKKGDILRIRDKPEEQWWNAEDSEGKRGMIPVPYVEKYRPASASVSALIGGNQEGSHPQPLGGPEPGPYAQPSVNTPLPNLQNGPIYARVIQKRVPNAYDKTALALEVGELVKVTKINVSGQWEGECNGKRGHFPFTHVRLLDQQNPDEDFS 4CFBFB65BFC2E265 paxi_human Paxillin PXN protein protein human Homo sapiens Human MDDLDALLADLESTTSHISKRPVFLSEETPYSYPTGNHTYQEIAVPPPVPPPPSSEALNGTILDPLDQWQPSSSRFIHQQPQSSSPVYGSSAKTSSVSNPQDSVGSPCSRVGEEEHVYSFPNKQKSAEPSPTVMSTSLGSNLSELDRLLLELNAVQHNPPGFPADEANSSPPLPGALSPLYGVPETNSPLGGKAGPLTKEKPKRNGGRGLEDVRPSVESLLDELESSVPSPVPAITVNQGEMSSPQRVTSTQQQTRISASSATRELDELMASLSDFKIQGLEQRADGERCWAAGWPRDGGRSSPGGQDEGGFMAQGKTGSSSPPGGPPKPGSQLDSMLGSLQSDLNKLGVATVAKGVCGACKKPIAGQVVTAMGKTWHPEHFVCTHCQEEIGSRNFFERDGQPYCEKDYHNLFSPRCYYCNGPILDKVVTALDRTWHPEHFFCAQCGAFFGPEGFHEKDGKAYCRKDYFDMFAPKCGGCARAILENYISALNTLWHPECFVCRECFTPFVNGSFFEHDGQPYCEVHYHERRGSLCSGCQKPITGRCITAMAKKFHPEHFVCAFCLKQLNKGTFKEQNDKPYCQNCFLKLFC DIP protein P49023 original sequence version: 115 DIP protein P49023 original sequence version: 126 DIP protein P49023 original sequence version: 169 DIP protein P49023 original sequence version: 140 DIP protein P49023 original sequence version: 175 ABF6C0BE5939623F fak1-pxn 2 5 unspecified role unspecified role bait bait Q05397:p.Val928Gly mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain V G Wed May 02 17:47:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val928gly' Tue Jun 30 17:17:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val928Gly' Q05397:p.Leu931Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L R Wed May 02 20:33:26 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu931arg' Tue Jun 30 20:15:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu931Arg' Q05397:p.Arg1042Gly mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R G Wed May 02 19:47:06 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1042gly' Tue Jun 30 16:12:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1042Gly' Q05397:p.Asp1039Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain D A Wed May 02 15:39:10 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp1039ala' Tue Jun 30 16:00:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp1039Ala' Q05397:p.Leu1035Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L A Wed May 02 18:03:17 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu1035ala' Tue Jun 30 18:06:28 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu1035Ala' Q05397:p.Val1037Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain V D Wed May 02 19:20:19 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val1037asp' Tue Jun 30 15:37:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val1037Asp' region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q05397:p.Val935Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain V A Wed May 02 16:07:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val935ala' Tue Jun 30 20:14:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val935Ala' Q05397:p.Leu1034Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L S Wed May 02 20:01:17 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu1034ser' Tue Jun 30 19:43:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu1034Ser' ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey human-hpb-all Homo sapiens hpb-all Human T lymphoblastoid cell line HPB-ALL This cell line has been shown to have been contaminated by Jurkat cells at some unknown point in its usage. physical association physical association Fig. 1A (Lane 8);Fig. 2A (Lane2,4); Fig. 5A,C; Fig 7A E.coli strain JM105 cells were used to produce GST-FAK fusion proteins. Paxillin proteins in this interaction were from a human T lymphoblastoid cell line, HPB-ALL. Cell lysates of HPB-ALL cells were precipitated with GST-FAK fusion proteins and then immunoblotted with anti-paxillin mAb. Cell lysates of HPB-ALL cells were incubated in uncoated (Fig.2A Lane 2) and FN-coated (Fig.2A Lane 4) plates. From the results, it is indicated that tyrosine phosphorylation is not required for the binding of Fak1 protein. Deletion analysis was conducted to determine the required region of Fak1 proteins for paxillin binding. (Fig.5 A,C ) Mutation analysis was conducted to determine the influence of specific amino acid mutations on the paxillin-bindng activity of Fak1. (Fig.7A) Cell lysates from the HPB-ALL cells in the FN-coated plates where paxillin got phosphorylated, were precipitated by the GST-FAK fusion proteins, and then immunoblotted with anti-phosphotyrosine mAb.(Fig1A, Lane8) fak1-pxn 2 5 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey human-t47d human breast cancer tumour cell line-epithelial cells t47d Epithelial cells derived from human breast tumour physical association physical association Fig. 2A(Lane 6),Fig. 2B(Lane 4,5) E.coli strain JM105 cells were used to produce GST-FAK fusion proteins. Paxillin proteins in this interaction were from a human breast cancer cell line, T-47D. Cell lysates of T-47D cells were precipitated with GST-FAK fusion protein and then immunoblotted with anti-paxillin mAb.(Fig2A, Lane 6) The pp70 was identified as paxillin by reprecipitating the pp70 (precipitated by GST-FAK) using anti-phopshotyrosine mAb, (Fig.2 Lane 4) anti-paxillin mAb (Fig.2 Lane 5) and immunoblotting with anti-paxilin mAb. fak1-pxn 3 5 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Fig. 3A (Lane 1,2) Cell lysates of HPB-ALL cells were precipitated with anti-paxillin mAb, and then analysed by immunoblotting with anti-FAK mAb. HPB-ALL cells were uncoated in Lane 1, and FN-coated in Lane 2. fak1-pxn 4 5 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Fig. 3A (Lane 3) Cell lysates of T-47D cells were precipitated with anti-paxillin mAb and then analysed by immunoblotting with anti-FAK mAb. fak1-crk 4 5 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association Fig. 3A (Lane5) Cell lysates of T-47D cells were precipitated with anti-crk mAb and then analysed by immunoblotting with anti-FAK mAb. fak1-pxn 1 5 unspecified role unspecified role bait bait region 125i radiolabel 125i radiolabel I125 125I 125I I125 autoradiography autoradiography undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey human-hpb-all Homo sapiens hpb-all Human T lymphoblastoid cell line HPB-ALL This cell line has been shown to have been contaminated by Jurkat cells at some unknown point in its usage. physical association physical association Fig.4A (Lane1) Paxillins were immunoprecipitated by anti-paxillin mAb from HPB-ALL cells, which were then analysed by overlay assay with 125i-labeled GST-FAK fusion proteins and autoradiographed. fak1-pxn 1 7 unspecified role unspecified role prey prey human-t47d human breast cancer tumour cell line-epithelial cells t47d Epithelial cells derived from human breast tumour 5 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 125i radiolabel 125i radiolabel I125 125I 125I I125 autoradiography autoradiography undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Fig. 4A (Lane2) Paxillins were immunoprecipitated by anti-paxillin mAb from T-47D cells, which were then analysed by overlay assay with 125i-labeled GST-FAK fusion proteins and autoradiographed.