mbinfo MBInfo http://www.mechanobio.info/ http://www.mechanobio.info/ rohatgi-2000-2 Mechanism of N-WASP activation by CDC42 and phosphatidylinositol 4, 5-bisphosphate. 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 J. Cell Biol. (0021-9525) 2000 Only protein-protein interactions imex curation rohatgi-2000-1 Mechanism of N-WASP activation by CDC42 and phosphatidylinositol 4, 5-bisphosphate. 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 J. Cell Biol. (0021-9525) 2000 Only protein-protein interactions imex curation arp2_bovin Actin-related protein 2 ACTR2 ARP2 Actin-like protein 2 protein protein bovin Bos taurus (Bovine) Bovine MDSQGRKVVVCDNGTGFVKCGYAGSNFPEHIFPALVGRPIIRSTTKVGNIEIKDLMVGDEASELRSMLEVNYPMENGIVRNWDDMKHLWDYTFGPEKLNIDTRNCKILLTEPPMNPTKNREKIVEVMFETYQFSGVYVAIQAVLTLYAQGLLTGVVVDSGDGVTHICPVYEGFSLPHLTRRLDIAGRDITRYLIKLLLLRGYAFNHSADFETVRMIKEKLCYVGYNIEQEQKLALETTVLVESYTLPDGRIIKVGGERFEAPEALFQPHLINVEGVGVAELLFNTIQAADIDTRSEFYKHIVLSGGSTMYPGLPSRLERELKQLYLERVLKGDVEKLSKFKIRIEDPPRRKHMVFLGGAVLADIMKDKDNFWMTRQEYQEKGVRVLEKLGVTVR wasl_bovin Neural Wiskott-Aldrich syndrome protein WASL protein protein bovin Bos taurus (Bovine) Bovine MSSGQQQPPPPRRVTNVGSLLLTPQENESLFTFLGKKCVTMSSAVVQLYAADRNCMWSKKCSGVACLVKDNPQRSYFLRIFDIKDGKLLWEQELYNNFVYNSPRGYFHTFAGDTCQVALNFANEEEAKKFRKAVTDLLGRRQRKSEKRRDPPNGPNLPMATVDIKNPEITTNRFYGPQINNISHTKEKKKGKAKKKRLTKADIGTPSNFQHIGHVGWDPNTGFDLNNLDPELKNLFDMCGISEAQLKDRETSKVIYDFIEKTGGVEAVKNELRRQAPPPPPPSRGGPPPPPPPPHSSGPPPPPARGRGAPPPPPSRAPTAAPPPPPPSRPGVGAPPPPPNRMYPPPLPALPSSAPSGPPPPPPPLSVSGSVAPPPPPPPPPPPGPPPPPGLPSDGDHQVPTPAGSKAALLDQIREGAQLKKVEQNSRPVSCSGRDALLDQIRQGIQLKSVTDAPESTPPAPAPTSGIVGALMEVMQKRSKAIHSSDEDEDEDDDEDFEDDDEWED arp3_bovin Actin-related protein 3 ACTR3 ARP3 Actin-like protein 3 Actin-2 protein protein bovin Bos taurus (Bovine) Bovine MAGRLPACVVDCGTGYTKLGYAGNTEPQFIIPSCIAIKESAKVGDQAQRRVMKGVDDLDFFIGDEAIEKPTYATKWPIRHGIVEDWDLMERFMEQVIFKYLRAEPEDHYFLLTEPPLNTPENREYTAEIMFESFNVPGLYIAVQAVLALAASWTSRQVGERTLTGTVIDSGDGVTHVIPVAEGYVIGSCIKHIPIAGRDITYFIQQLLRDREVGIPPEQSLETAKAVKERYSYVCPDLVKEFNKYDTDGSKWIKQYTGINAISKKEFSIDVGYERFLGPEIFFHPEFANPDFTQPISEVVDEVIQNCPIDVRRPLYKNIVLSGGSTMFRDFGRRLQRDLKRTVDARLKLSEELSGGRLKPKPIDVQVITHHMQRYAVWFGGSMLASTPEFYQVCHTKKDYEEIGPSICRHNPVFGVMS 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 wasl-actr2 1316777 n/a 1316779 unspecified role unspecified role prey prey bovin-brain Bos taurus brain brain Brain [cerebrum] n/a 1316780 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 false vca 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) physical association physical association false false false Fig 2B. wasl-actr3-3 1316777 n/a 1316781 unspecified role unspecified role prey prey bovin-brain Bos taurus brain brain Brain [cerebrum] n/a 1316782 unspecified role unspecified role prey prey bovin-brain Bos taurus brain brain Brain [cerebrum] n/a 1316780 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 false vca 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) association association false false false 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 1316777 n/a 1316780 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 false wg fragment sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 n/a 1316780 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 false 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 false ecoli Escherichia coli (strain K12) 1316799 1316797 association association false false false Fig 2C, 2D. wasl-actr3-4 1316777 n/a 1316781 unspecified role unspecified role prey prey bovin-brain Bos taurus brain brain Brain [cerebrum] n/a 1316780 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 false 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 false ecoli Escherichia coli (strain K12) association association false false false 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. wasl-wasl-4 1316778 n/a 1316780 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 false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false 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 false rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes n/a 1316780 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 false 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 false ecoli Escherichia coli (strain K12) 1316812 1316810 physical association physical association false false false Fig 1B. wasl-wasl-6 1316778 n/a 1316780 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 false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false wg fragment sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes n/a 1316780 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 false 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 false ecoli Escherichia coli (strain K12) 1316820 1316818 physical association physical association false false false Fig 1D.