mbinfo MBInfo http://www.mechanobio.info/ Integration of multiple signals through cooperative regulation of the N-WASP-Arp2/3 complex. Integration of multiple signals through cooperative regulation of the N-WASP-Arp2/3 complex. Science (0036-8075) 2000 imex curation Prehoda K.E., Scott J.A., Mullins R.D., Lim W.A. Interaction detection: GST pull down. The source of N-WASP specified by the author. wlim@itsa.ucsf.edu 11-Oct-2012: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro cosedimentation cosedimentation weight by comassie molecular weight estimation by coomasie staining weight by comassie Prehoda K.E., Scott J.A., Mullins R.D., Lim W.A. Interaction detection: GST pull down. The source of N-WASP specified by the author. wlim@itsa.ucsf.edu Accepted 2012-SEP-20 by ORCHARD 11-Oct-2012: Contacted by IntAct-Help. Science (0036-8075) 2000 Only protein-protein interactions imex curation Integration of multiple signals through cooperative regulation of the N-WASP-Arp2/3 complex. Integration of multiple signals through cooperative regulation of the N-WASP-Arp2/3 complex. Science (0036-8075) 2000 imex curation Prehoda K.E., Scott J.A., Mullins R.D., Lim W.A. Interaction detection: GST pull down. The source of N-WASP specified by the author. wlim@itsa.ucsf.edu 11-Oct-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 Prehoda K.E., Scott J.A., Mullins R.D., Lim W.A. Interaction detection: GST pull down. The source of N-WASP specified by the author. The source of N-WASP specified by the author. wlim@itsa.ucsf.edu Accepted 2012-SEP-20 by ORCHARD 11-Oct-2012: Contacted by IntAct-Help. Science (0036-8075) 2000 Only protein-protein interactions imex curation Integration of multiple signals through cooperative regulation of the N-WASP-Arp2/3 complex. Integration of multiple signals through cooperative regulation of the N-WASP-Arp2/3 complex. Science (0036-8075) 2000 imex curation Prehoda K.E., Scott J.A., Mullins R.D., Lim W.A. Interaction detection: GST pull down. The source of N-WASP specified by the author. wlim@itsa.ucsf.edu 11-Oct-2012: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Prehoda K.E., Scott J.A., Mullins R.D., Lim W.A. Interaction detection: GST pull down. The source of N-WASP specified by the author. wlim@itsa.ucsf.edu Accepted 2012-SEP-20 by ORCHARD 11-Oct-2012: Contacted by IntAct-Help. Science (0036-8075) 2000 Only protein-protein interactions imex curation 1-phosphatidyl-1d-myo-inositol 4,5-bisphosphate 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) wasl_rat Neural Wiskott-Aldrich syndrome protein Wasl protein protein rat Rattus norvegicus (Rat) Rat MSSGQQPPRRVTNVGSLLLTPQENESLFSFLGKKCVTMSSAVVQLYAADRNCMWSKKCSGVACLVKDNPQRSYFLRIFDIKDGKLLWEQELYNNFVYNSPRGYFHTFAGDTCQVALNFANEEEAKKFRKAVTDLLGRRQRKSEKRRDAPNGPNLPMATVDIKNPEITTNRFYSSQVNNISHTKEKKKGKAKKKRLTKADIGTPSNFQHIGHVGWDPNTGFDLNNLDPELKNLFDMCGISEAQLKDRETSKVIYDFIEKTGGVEAVKNELRRQAPPPPPPSRGGPPPPPPPPHSSGPPPPPARGRGAPPPPPSRAPTAAPPPPPPSRPGVVVPPPPPNRMYPPPPPALPSSAPSGPPPPPPLSMAGSTAPPPPPPPPPPPGPPPPPGLPSDGDHQVPASSGNKAALLDQIREGAQLKKVEQNSRPVSCSGRDALLDQIRQGIQLKSVSDGQESTPPTPAPTSGIVGALMEVMQKRSKAIHSSDEDEDDDDEEDFQDDDEWED DIP protein O08816 original sequence version: 145 DIP protein O08816 original sequence version: 115 480E21F26F7FC77E arp2_acaca Actin-related protein 2 arp2 Actin-like protein 2 protein protein acaca Acanthamoeba castellanii (Amoeba) MTDSSKVIVCDNGTGFVKCGFARSNFPASIFPSMVGRPILRSEEKFDNVEIKDIMVGDEASKLRSMLQITYPLDNGIVRNWEDAEHVWNYTFFEKLKVDPKDCKILLTEPPMNPLANREKMVQVMFEKYGFKAAYIAIQAVLTLYAQGLLTGVVVDSGDGVTHIVPVYEGFSLPHLTRRLNVAGRDVTRYLIKLLLLRGYVFNRTADFETVRQIKEKFCYVGYDLELEKRLALETTTLVEKYTLPDGRVIPIGAERFEAPECMFNPALVDQESVGVGELVFDCINKADIDTRAEFYNHIVLSGGSTMYPGLPSRLEKEIKRLYFERVAKGNKVSMQKFKCRIEDPPRRKHMVFLGGAVLAEIMKDKTAFWMNKSEYEEQGPRVLRKCF 456C31CDC70B4F70 cdc42_human Cell division control protein 42 homolog CDC42 G25K GTP-binding protein protein protein human Homo sapiens Human MQTIKCVVVGDGAVGKTCLLISYTTNKFPSEYVPTVFDNYAVTVMIGGEPYTLGLFDTAGQEDYDRLRPLSYPQTDVFLVCFSVVSPSSFENVKEKWVPEITHHCPKTPFLLVGTQIDLRDDPSTIEKLAKNKQKPITPETAEKLARDLKAVKYVECSALTQKGLKNVFDEAILAALEPPEPKKSRRCVLL 51A437E22A4D8FFF wasl-wasl-2 3 5 unspecified role unspecified role prey prey vca region sufficient to bind sufficient binding region sufficient to bind 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 ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role bait bait control region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain The control region contains within it the basic region and the GTPase-binding domain of N-WASP. 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) 10 13 direct interaction direct interaction Fig 1B (Bottom panel). wasl-arp2 2 6 unspecified role unspecified role prey prey acaca Acanthamoeba castellanii (Amoeba) 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 control region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain The control region contains within it both the basic region and the GTPase-binding domain. ecoli Escherichia coli (strain K12) physical association physical association Fig 2A(right panel), 2B. The interaction between the N-WASP-control region and Arp2 was inhibited in the presence of increasing concentrations of Cdc42-GTPgammaS. 1_phosphatidyl-cdc42 1 7 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey control region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain The control region contains within it the basic region and the GTPase-binding domain of N-WASP. ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role bait bait physical association physical association Fig 3B. N-WASP and Cdc42 get cosedimented to the pellet due to the presence of PIP2 vesicles, which bind simultaneously to N-WASP.