mbinfo MBInfo http://www.mechanobio.info/ http://www.mechanobio.info/ uruno-2003-1 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. 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 J. Biol. Chem. (0021-9258) 2003 Only protein-protein interactions imex curation src8_mouse Src substrate cortactin Cttn Ems1 protein protein mouse Mus musculus Mouse MWKASAGHAVSITQDDGGADDWETDPDFVNDVSEKEQRWGAKTVQGSGHQEHINIHKLRENVFQEHQTLKEKELETGPKASHGYGGKFGVEQDRMDRSAVGHEYQSKLSKHCSQVDSVRGFGGKFGVQMDRVDQSAVGFEYQGKTEKHASQKDYSSGFGGKYGVQADRVDKSAVGFDYQGKTEKHESQKDYSKGFGGKYGIDKDKVDKSAVGFEYQGKTEKHESQKDYVKGFGGKFGVQTDRQDKCALGWDHQEKLQLHESQKDYKTGFGGKFGVQSERQDSSAVGFDYKERLAKHESQQDYAKGFGGKYGVQKDRMDKNASTFEEVVQVPSAYQKTVPIEAVTSKTSNIRANFENLAKEREQEDRRKAEAERAQRMAKERQEQEEARRKLEEQARAKKQTPPASPSPQPIEDRPPSSPIYEDAAPFKAEPSYRGSEPEPEYSIEAAGIPEAGSQQGLTYTSEPVYETTEAPGHYQAEDDTYDGYESDLGITAIALYDYQAAGDDEISFDPDDIITNIEMIDDGWWRGVCKGRYGLFPANYVELRQ arp3_bovin Actin-related protein 3 ACTR3 ARP3 Actin-like protein 3 Actin-2 protein protein bovin Bos taurus (Bovine) Bovine MAGRLPACVVDCGTGYTKLGYAGNTEPQFIIPSCIAIKESAKVGDQAQRRVMKGVDDLDFFIGDEAIEKPTYATKWPIRHGIVEDWDLMERFMEQVIFKYLRAEPEDHYFLLTEPPLNTPENREYTAEIMFESFNVPGLYIAVQAVLALAASWTSRQVGERTLTGTVIDSGDGVTHVIPVAEGYVIGSCIKHIPIAGRDITYFIQQLLRDREVGIPPEQSLETAKAVKERYSYVCPDLVKEFNKYDTDGSKWIKQYTGINAISKKEFSIDVGYERFLGPEIFFHPEFANPDFTQPISEVVDEVIQNCPIDVRRPLYKNIVLSGGSTMFRDFGRRLQRDLKRTVDARLKLSEELSGGRLKPKPIDVQVITHHMQRYAVWFGGSMLASTPEFYQVCHTKKDYEEIGPSICRHNPVFGVMS wasl_human Neural Wiskott-Aldrich syndrome protein WASL protein protein human Homo sapiens Human MSSVQQQPPPPRRVTNVGSLLLTPQENESLFTFLGKKCVTMSSAVVQLYAADRNCMWSKKCSGVACLVKDNPQRSYFLRIFDIKDGKLLWEQELYNNFVYNSPRGYFHTFAGDTCQVALNFANEEEAKKFRKAVTDLLGRRQRKSEKRRDPPNGPNLPMATVDIKNPEITTNRFYGPQVNNISHTKEKKKGKAKKKRLTKADIGTPSNFQHIGHVGWDPNTGFDLNNLDPELKNLFDMCGISEAQLKDRETSKVIYDFIEKTGGVEAVKNELRRQAPPPPPPSRGGPPPPPPPPHNSGPPPPPARGRGAPPPPPSRAPTAAPPPPPPSRPSVAVPPPPPNRMYPPPPPALPSSAPSGPPPPPPSVLGVGPVAPPPPPPPPPPPGPPPPPGLPSDGDHQVPTTAGNKAALLDQIREGAQLKKVEQNSRPVSCSGRDALLDQIRQGIQLKSVADGQESTPPTPAPTSGIVGALMEVMQKRSKAIHSSDEDEDEDDEEDFEDDDEWED cttn-actr3-10 1311932 n/a 1311933 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 false n-terminal region 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 1311934 unspecified role unspecified role prey prey bovin Bos taurus (Bovine) Bovine physical association physical association false false false Fig 1. Binding of Arp3 to cortactin was inhibited in the presence of increasing concentrations of N-WASP-VCA domain. wasl-actr3-2 1311932 n/a 1311935 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 false vca region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false The amino acid range for the VCA region traced from references [PMID:10491394]. ecoli Escherichia coli (strain K12) n/a 1311934 unspecified role unspecified role prey prey bovin Bos taurus (Bovine) Bovine physical association physical association false false false 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).