mbinfo MBInfo http://www.mechanobio.info/ Mechanism of N-WASP activation by CDC42 and phosphatidylinositol 4, 5-bisphosphate. Mechanism of N-WASP activation by CDC42 and phosphatidylinositol 4, 5-bisphosphate. J. Cell Biol. (0021-9525) 2000 imex curation Rohatgi R., Ho H.Y., Kirschner M.W. Interaction detection: GST pull down. Interactions with Cdc42 (Fig 2E) could not be curated since its source could not be traced. Interactions with unlabeled G-actin (Fig 3) could not be curated since its source could not be clearly traced. marc@hms.harvard.edu 11-Oct-2012: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down autoradiography autoradiography Rohatgi R., Ho H.Y., Kirschner M.W. Interaction detection: GST pull down. Interactions with Cdc42 (Fig 2E) could not be curated since its source could not be traced. Interactions with unlabeled G-actin (Fig 3) could not be curated since its source could not be clearly traced. marc@hms.harvard.edu Accepted 2012-SEP-21 by ORCHARD 11-Oct-2012: Contacted by IntAct-Help. J. Cell Biol. (0021-9525) 2000 Only protein-protein interactions imex curation Mechanism of N-WASP activation by CDC42 and phosphatidylinositol 4, 5-bisphosphate. Mechanism of N-WASP activation by CDC42 and phosphatidylinositol 4, 5-bisphosphate. J. Cell Biol. (0021-9525) 2000 imex curation Rohatgi R., Ho H.Y., Kirschner M.W. Interaction detection: GST pull down. Interactions with Cdc42 (Fig 2E) could not be curated since its source could not be traced. Interactions with unlabeled G-actin (Fig 3) could not be curated since its source could not be clearly traced. marc@hms.harvard.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 Rohatgi R., Ho H.Y., Kirschner M.W. Interaction detection: GST pull down. Interactions with Cdc42 (Fig 2E) could not be curated since its source could not be traced. Interactions with unlabeled G-actin (Fig 3) could not be curated since its source could not be clearly traced. marc@hms.harvard.edu Accepted 2012-SEP-21 by ORCHARD 11-Oct-2012: Contacted by IntAct-Help. J. Cell Biol. (0021-9525) 2000 Only protein-protein interactions imex curation 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 arp2_bovin Actin-related protein 2 ACTR2 ARP2 Actin-like protein 2 protein protein bovin Bos taurus (Bovine) Bovine MDSQGRKVVVCDNGTGFVKCGYAGSNFPEHIFPALVGRPIIRSTTKVGNIEIKDLMVGDEASELRSMLEVNYPMENGIVRNWDDMKHLWDYTFGPEKLNIDTRNCKILLTEPPMNPTKNREKIVEVMFETYQFSGVYVAIQAVLTLYAQGLLTGVVVDSGDGVTHICPVYEGFSLPHLTRRLDIAGRDITRYLIKLLLLRGYAFNHSADFETVRMIKEKLCYVGYNIEQEQKLALETTVLVESYTLPDGRIIKVGGERFEAPEALFQPHLINVEGVGVAELLFNTIQAADIDTRSEFYKHIVLSGGSTMYPGLPSRLERELKQLYLERVLKGDVEKLSKFKIRIEDPPRRKHMVFLGGAVLADIMKDKDNFWMTRQEYQEKGVRVLEKLGVTVR DIP protein A7MB62 original sequence version: 6 DIP protein A7MB62 original sequence version: 35 DIP protein A7MB62 original sequence version: 47 1BFA6B442ED1A797 wasl_bovin Neural Wiskott-Aldrich syndrome protein WASL protein protein bovin Bos taurus (Bovine) Bovine MSSGQQQPPPPRRVTNVGSLLLTPQENESLFTFLGKKCVTMSSAVVQLYAADRNCMWSKKCSGVACLVKDNPQRSYFLRIFDIKDGKLLWEQELYNNFVYNSPRGYFHTFAGDTCQVALNFANEEEAKKFRKAVTDLLGRRQRKSEKRRDPPNGPNLPMATVDIKNPEITTNRFYGPQINNISHTKEKKKGKAKKKRLTKADIGTPSNFQHIGHVGWDPNTGFDLNNLDPELKNLFDMCGISEAQLKDRETSKVIYDFIEKTGGVEAVKNELRRQAPPPPPPSRGGPPPPPPPPHSSGPPPPPARGRGAPPPPPSRAPTAAPPPPPPSRPGVGAPPPPPNRMYPPPLPALPSSAPSGPPPPPPPLSVSGSVAPPPPPPPPPPPGPPPPPGLPSDGDHQVPTPAGSKAALLDQIREGAQLKKVEQNSRPVSCSGRDALLDQIRQGIQLKSVTDAPESTPPAPAPTSGIVGALMEVMQKRSKAIHSSDEDEDEDDDEDFEDDDEWED 54B83B48F1CDB3B8 arpc2_bovin Actin-related protein 2/3 complex subunit 2 ARPC2 Arp2/3 complex 34 kDa subunit protein protein bovin Bos taurus (Bovine) Bovine MILLEVNNRIIEETLALKFENAAAGNKPEAVEVTFADFDGVLYHISNPNGDKTKVMVSISLKFYKELQAHGADELLKRVYGSYLVNPESGYNVSLLYDLENLPASKDSIVHQAGMLKRNCFASVFEKYFQFQEEGKEGENRAVIHYRDDETMYVESKKDRVTVVFSTVFKDDDDVVIGKVFMQEFKEGRRASHTAPQVLFSHREPPLELKDTDAAVGDNIGYITFVLFPRHTNASARDNTINLIHTFRDYLHYHIKCSKAYIHTRMRAKTSDFLKVLNRARPDAEKKEMKTITGKTFSSR DIP protein Q3MHR7 original sequence version: 23 DIP protein Q3MHR7 original sequence version: 52 DIP protein Q3MHR7 original sequence version: 63 60801D999BFBF947 wasl-wasl-4 1 5 unspecified role unspecified role prey prey n-terminal region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 35s-methionine labelled Myc-N-WASP. gbr fragment sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain The GBR fragment contains within it both the Basic region as well as the GTPase-Binding Domain (GBD). rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes 5 unspecified role unspecified role bait bait vca region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 11 13 physical association physical association Fig 1B. wasl-wasl-6 1 5 unspecified role unspecified role prey prey n-terminal region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 35s-methionine-labeled Myc-N-WASP. wg fragment sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain The WG fragment contains within it the WH1 domain, the basic region and the GTPase-binding domain. rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes 5 unspecified role unspecified role bait bait ca region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain The CA region contains within the Cofilin homology region and the acidic domain. n-terminal 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) 19 21 physical association physical association Fig 1D. wasl-actr2 2 4 unspecified role unspecified role prey prey bovin-brain Bos taurus brain brain Brain [cerebrum] 5 unspecified role unspecified role bait bait n-terminal region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range vca 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) physical association physical association Fig 2B. wasl-actr3-3 2 3 unspecified role unspecified role prey prey bovin-brain Bos taurus brain brain Brain [cerebrum] 6 unspecified role unspecified role prey prey bovin-brain Bos taurus brain brain Brain [cerebrum] 5 unspecified role unspecified role bait bait n-terminal region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range vca 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) association association Fig 2C. The association of the Arp2/3 complex subunits with N-WASP-VCA was inhibited in the presence of increasing concentrations of N-WASP-WG and N-WASP-WGP fragments. wasl-wasl-8 2 5 unspecified role unspecified role prey prey n-terminal region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range wg fragment sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain The WG fragment contains within it the WH1 domain, the basic region and the GTPase-binding domain. spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 5 unspecified role unspecified role bait bait vca region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 37 39 association association Fig 2C, 2D. wasl-actr3-4 2 3 unspecified role unspecified role prey prey bovin-brain Bos taurus brain brain Brain [cerebrum] 5 unspecified role unspecified role bait bait vca region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) association association Fig 2D. The interaction was inhibited in the presence of increasing concentrations of N-WASP-WG, but this inhibition was removed by stimulation with Cdc42 and PIP2.