mbinfo MBInfo http://www.mechanobio.info/ Sequential interaction of actin-related proteins 2 and 3 (Arp2/3) complex with neural Wiscott-Aldrich syndrome protein (N-WASP) and cortactin during branched actin filament network formation. Sequential interaction of actin-related proteins 2 and 3 (Arp2/3) complex with neural Wiscott-Aldrich syndrome protein (N-WASP) and cortactin during branched actin filament network formation. J. Biol. Chem. (0021-9258) 2003 imex curation Uruno T., Liu J., Li Y., Smith N., Zhan X. Interaction detection method: GST pull down. The amount of Arp3 pulled down by the bead-bound cortactin was quantitated. The source of cortactin and Arp3 specified by the author. zhanx@usa.redcross.org 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 Uruno T., Liu J., Li Y., Smith N., Zhan X. Interaction detection method: GST pull down. The amount of Arp3 pulled down by the bead-bound cortactin was quantitated. The source of cortactin and Arp3 specified by the author. zhanx@usa.redcross.org Accepted 2012-AUG-08 by ORCHARD 19-Sept-2012: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2003 Only protein-protein interactions imex curation wasl_human Neural Wiskott-Aldrich syndrome protein WASL protein protein human Homo sapiens Human MSSVQQQPPPPRRVTNVGSLLLTPQENESLFTFLGKKCVTMSSAVVQLYAADRNCMWSKKCSGVACLVKDNPQRSYFLRIFDIKDGKLLWEQELYNNFVYNSPRGYFHTFAGDTCQVALNFANEEEAKKFRKAVTDLLGRRQRKSEKRRDPPNGPNLPMATVDIKNPEITTNRFYGPQVNNISHTKEKKKGKAKKKRLTKADIGTPSNFQHIGHVGWDPNTGFDLNNLDPELKNLFDMCGISEAQLKDRETSKVIYDFIEKTGGVEAVKNELRRQAPPPPPPSRGGPPPPPPPPHNSGPPPPPARGRGAPPPPPSRAPTAAPPPPPPSRPSVAVPPPPPNRMYPPPPPALPSSAPSGPPPPPPSVLGVGPVAPPPPPPPPPPPGPPPPPGLPSDGDHQVPTTAGNKAALLDQIREGAQLKKVEQNSRPVSCSGRDALLDQIRQGIQLKSVADGQESTPPTPAPTSGIVGALMEVMQKRSKAIHSSDEDEDEDDEEDFEDDDEWED DIP protein O00401 original sequence version: 55 DIP protein O00401 original sequence version: 147 DIP protein O00401 original sequence version: 150 DIP protein O00401 original sequence version: 136 DIP protein O00401 original sequence version: 138 DIP protein O00401 original sequence version: 125 DIP protein O00401 original sequence version: 112 DIP protein O00401 original sequence version: 126 B5895C6055E23049 src8_mouse Src substrate cortactin Cttn Ems1 protein protein mouse Mus musculus Mouse MWKASAGHAVSITQDDGGADDWETDPDFVNDVSEKEQRWGAKTVQGSGHQEHINIHKLRENVFQEHQTLKEKELETGPKASHGYGGKFGVEQDRMDRSAVGHEYQSKLSKHCSQVDSVRGFGGKFGVQMDRVDQSAVGFEYQGKTEKHASQKDYSSGFGGKYGVQADRVDKSAVGFDYQGKTEKHESQKDYSKGFGGKYGIDKDKVDKSAVGFEYQGKTEKHESQKDYVKGFGGKFGVQTDRQDKCALGWDHQEKLQLHESQKDYKTGFGGKFGVQSERQDSSAVGFDYKERLAKHESQQDYAKGFGGKYGVQKDRMDKNASTFEEVVQVPSAYQKTVPIEAVTSKTSNIRANFENLAKEREQEDRRKAEAERAQRMAKERQEQEEARRKLEEQARAKKQTPPASPSPQPIEDRPPSSPIYEDAAPFKAEPSYRGSEPEPEYSIEAAGIPEAGSQQGLTYTSEPVYETTEAPGHYQAEDDTYDGYESDLGITAIALYDYQAAGDDEISFDPDDIITNIEMIDDGWWRGVCKGRYGLFPANYVELRQ DIP protein Q60598 original sequence version: 136 DIP protein Q60598 original sequence version: 78 DIP protein Q60598 original sequence version: 135 8F5CA026ACCD6D4F arp3_bovin Actin-related protein 3 ACTR3 ARP3 Actin-like protein 3 Actin-2 protein protein bovin Bos taurus (Bovine) Bovine MAGRLPACVVDCGTGYTKLGYAGNTEPQFIIPSCIAIKESAKVGDQAQRRVMKGVDDLDFFIGDEAIEKPTYATKWPIRHGIVEDWDLMERFMEQVIFKYLRAEPEDHYFLLTEPPLNTPENREYTAEIMFESFNVPGLYIAVQAVLALAASWTSRQVGERTLTGTVIDSGDGVTHVIPVAEGYVIGSCIKHIPIAGRDITYFIQQLLRDREVGIPPEQSLETAKAVKERYSYVCPDLVKEFNKYDTDGSKWIKQYTGINAISKKEFSIDVGYERFLGPEIFFHPEFANPDFTQPISEVVDEVIQNCPIDVRRPLYKNIVLSGGSTMFRDFGRRLQRDLKRTVDARLKLSEELSGGRLKPKPIDVQVITHHMQRYAVWFGGSMLASTPEFYQVCHTKKDYEEIGPSICRHNPVFGVMS DIP protein P61157 original sequence version: 44 DIP protein P61157 original sequence version: 74 DIP protein P61157 original sequence version: 86 23E5564198B81C63 cttn-actr3-10 1 3 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 n-terminal 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) 4 unspecified role unspecified role prey prey bovin Bos taurus (Bovine) Bovine physical association physical association Fig 1. Binding of Arp3 to cortactin was inhibited in the presence of increasing concentrations of N-WASP-VCA domain. wasl-actr3-2 1 2 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 vca region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain The amino acid range for the VCA region traced from references [PMID:10491394]. ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role prey prey bovin Bos taurus (Bovine) Bovine physical association physical association Fig 4A, 4B, 6A, 6B and 6C. The Arp3 immunoblot detects not the N-WASP-VCA-bound fraction from the pellet, but the unbound fraction from the supernatant. Thus, the band intensity is inversely proportional to the amount of Arp3 bound to GST-VCA. The binding of Arp3 to N-WASP-VCA was inhibited in the presence of G-actin and increasing concentrations of cortactin. In the absence of G-actin, increasing concentrations of cortactin had no effect on the Arp3-VCA interaction (Fig 4A, 4B). The Arp3-N-WASP-VCA interaction was proportional to the amount of debranched actin associated with Arp3, since the presence of branched actin associated with Arp3 inhibited the interaction (Fig 6A, 6B). The presence of cortactin could not completely disrupt the interaction, since cortactin has a lower affinity for F-actin-free Arp3 than N-WASP-VCA (Fig 6C).