mbinfo MBInfo http://www.mechanobio.info/ http://www.mechanobio.info/ cory-2002-3 Phosphorylation of tyrosine 291 enhances the ability of WASp to stimulate actin polymerization and filopodium formation. Wiskott-Aldrich Syndrome protein. in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Cory G.O., Garg R., Cramer R., Ridley A.J. Coimmunoprecipitations of Cdc42 with WASp (Fig 1D) could not be curated since the source of Cdc42 could not be identified. anne@ludwig. ucl.ac.uk J. Biol. Chem. (0021-9258) 2002 Only protein-protein interactions imex curation cory-2002-1 Phosphorylation of tyrosine 291 enhances the ability of WASp to stimulate actin polymerization and filopodium formation. Wiskott-Aldrich Syndrome protein. chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Cory G.O., Garg R., Cramer R., Ridley A.J. Coimmunoprecipitations of Cdc42 with WASp (Fig 1D) could not be curated since the source of Cdc42 could not be identified. anne@ludwig. ucl.ac.uk J. Biol. Chem. (0021-9258) 2002 Only protein-protein interactions imex curation cory-2002-2 Phosphorylation of tyrosine 291 enhances the ability of WASp to stimulate actin polymerization and filopodium formation. Wiskott-Aldrich Syndrome protein. chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Cory G.O., Garg R., Cramer R., Ridley A.J. The phosphorylated form of Hck, coimmunoprecipitated with WASp was detected by immunoblotting with anti-phosphotyrosine antibody 4G10. Coimmunoprecipitations of Cdc42 with WASp (Fig 1D) could not be curated since the source of Cdc42 could not be identified. anne@ludwig. ucl.ac.uk J. Biol. Chem. (0021-9258) 2002 Only protein-protein interactions imex curation wasp_human Wiskott-Aldrich syndrome protein WAS IMD2 protein protein human Homo sapiens Human MSGGPMGGRPGGRGAPAVQQNIPSTLLQDHENQRLFEMLGRKCLTLATAVVQLYLALPPGAEHWTKEHCGAVCFVKDNPQKSYFIRLYGLQAGRLLWEQELYSQLVYSTPTPFFHTFAGDDCQAGLNFADEDEAQAFRALVQEKIQKRNQRQSGDRRQLPPPPTPANEERRGGLPPLPLHPGGDQGGPPVGPLSLGLATVDIQNPDITSSRYRGLPAPGPSPADKKRSGKKKISKADIGAPSGFKHVSHVGWDPQNGFDVNNLDPDLRSLFSRAGISEAQLTDAETSKLIYDFIEDQGGLEAVRQEMRRQEPLPPPPPPSRGGNQLPRPPIVGGNKGRSGPLPPVPLGIAPPPPTPRGPPPPGRGGPPPPPPPATGRSGPLPPPPPGAGGPPMPPPPPPPPPPPSSGNGPAPPPLPPALVPAGGLAPGGGRGALLDQIRQGIQLNKTPGAPESSALQPPPQSSEGLVGALMHVMQKRSRAIHSSDEGEDQAGDEDEDDEWDD hck_mouse Tyrosine-protein kinase HCK Hck B-cell/myeloid kinase Hematopoietic cell kinase Hemopoietic cell kinase p56-HCK/p59-HCK protein protein mouse Mus musculus Mouse MGGRSSCEDPGCPRSEGRAPRMGCVKSRFLRDGSKASKTEPSANQKGPVYVPDPTSSSKLGPNNSNSMPPGFVEGSEDTIVVALYDYEAIHREDLSFQKGDQMVVLEEAGEWWKARSLATKKEGYIPSNYVARVNSLETEEWFFKGISRKDAERHLLAPGNMLGSFMIRDSETTKGSYSLSVRDFDPQHGDTVKHYKIRTLDSGGFYISPRSTFSSLQELVLHYKKGKDGLCQKLSVPCVSPKPQKPWEKDAWEIPRESLQMEKKLGAGQFGEVWMATYNKHTKVAVKTMKPGSMSVEAFLAEANLMKSLQHDKLVKLHAVVSQEPIFIVTEFMAKGSLLDFLKSEEGSKQPLPKLIDFSAQISEGMAFIEQRNYIHRDLRAANILVSASLVCKIADFGLARIIEDNEYTAREGAKFPIKWTAPEAINFGSFTIKSDVWSFGILLMEIVTYGRIPYPGMSNPEVIRALEHGYRMPRPDNCPEELYNIMIRCWKNRPEERPTFEYIQSVLDDFYTATESQYQQQP hck-was-7 1354013 n/a 1354016 enzyme target enzyme target substrate substrate unspecified role unspecified role 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 false crib domain 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 1354017 enzyme enzyme unspecified role unspecified role region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 phosphorylation phosphorylation reaction phosphorylation false false false Fig 3B. hck-1 1354013 n/a 1354017 putative self putative self putative self putative self n-terminal region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 phosphorylation phosphorylation reaction phosphorylation false true false Fig 3B. Interaction type: Autophosphorylation of Hck. was-hck-7 1354014 n/a 1354016 unspecified role unspecified role bait bait c-terminal region vsv tag vsv tag vesicular stomatitis virus tag vesicular stomatitis virus tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1354017 unspecified role unspecified role prey prey n-terminal region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Fig 1C. The interaction between WASp and Hck was unaffected in the presence of the kinase dead mutant of Hck (267 Hck). The interaction was also unaffected by the presence or absence of active (V12) or dominant negative (N17) CDC42. was-hck-9 1354015 n/a 1354017 unspecified role unspecified role prey prey n-terminal region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1354016 unspecified role unspecified role bait bait c-terminal region vsv tag vsv tag vesicular stomatitis virus tag vesicular stomatitis virus tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Fig 2A. Phospho forms of Hck were not detected by the anti-phosphotyrosine antibodies when the kinase dead mutant Hck (267 Hck) were coimmunoprecipitated with WASp, showing an absence of phosphorylation by the kinase dead mutant (Fig 1B). Phospo forms of Hck were not detected when Hck was coimmunoprecipitated with WASp in the presence of phosphatases such as CIP, PP1 and LAR (Fig 2A).