IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ Coronin 1B coordinates Arp2/3 complex and cofilin activities at the leading edge. Coronin 1B coordinates Arp2/3 complex and cofilin activities at the leading edge. Cell (0092-8674) 2007 imex curation Cai L., Marshall TW., Uetrecht AC., Schafer DA., Bear JE. jbear@email.unc.edu 26-APR-2007: Contacted by JYOTI. orchard Only protein-protein interactions 7 rat-rat-2 Rat R2 embryo cells rat-2 Rat embryo cell line anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Cai L., Marshall TW., Uetrecht AC., Schafer DA., Bear JE. jbear@email.unc.edu Accepted 2007-MAY-5 by ORCHARD. 26-APR-2007: Contacted by JYOTI. Cell (0092-8674) 2007 orchard Only protein-protein interactions 7 imex curation Coronin 1B coordinates Arp2/3 complex and cofilin activities at the leading edge. Coronin 1B coordinates Arp2/3 complex and cofilin activities at the leading edge. Cell (0092-8674) 2007 imex curation Cai L., Marshall TW., Uetrecht AC., Schafer DA., Bear JE. jbear@email.unc.edu 26-APR-2007: Contacted by JYOTI. orchard Only protein-protein interactions 7 in vitro In vitro pull down pull down predetermined predetermined participant predetermined Cai L., Marshall TW., Uetrecht AC., Schafer DA., Bear JE. It is stated in the reference given by the authors (Biochemistry 1999 38 15212-15222) that Arp2/3 was purified from human leukocytes. jbear@email.unc.edu Accepted 2007-MAY-5 by ORCHARD. 26-APR-2007: Contacted by JYOTI. Cell (0092-8674) 2007 orchard Only protein-protein interactions 7 imex curation Coronin 1B coordinates Arp2/3 complex and cofilin activities at the leading edge. Coronin 1B coordinates Arp2/3 complex and cofilin activities at the leading edge. Cell (0092-8674) 2007 imex curation Cai L., Marshall TW., Uetrecht AC., Schafer DA., Bear JE. jbear@email.unc.edu 26-APR-2007: Contacted by JYOTI. orchard Only protein-protein interactions 7 rat-rat-2 Rat R2 embryo cells rat-2 Rat embryo cell line anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Cai L., Marshall TW., Uetrecht AC., Schafer DA., Bear JE. jbear@email.unc.edu Accepted 2007-MAY-5 by ORCHARD. 26-APR-2007: Contacted by JYOTI. Cell (0092-8674) 2007 orchard Only protein-protein interactions 7 imex curation Coronin 1B coordinates Arp2/3 complex and cofilin activities at the leading edge. Coronin 1B coordinates Arp2/3 complex and cofilin activities at the leading edge. Cell (0092-8674) 2007 imex curation Cai L., Marshall TW., Uetrecht AC., Schafer DA., Bear JE. jbear@email.unc.edu 26-APR-2007: Contacted by JYOTI. orchard Only protein-protein interactions 7 human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Cai L., Marshall TW., Uetrecht AC., Schafer DA., Bear JE. jbear@email.unc.edu Accepted 2007-MAY-5 by ORCHARD. 26-APR-2007: Contacted by JYOTI. Cell (0092-8674) 2007 orchard Only protein-protein interactions 7 imex curation Coronin 1B coordinates Arp2/3 complex and cofilin activities at the leading edge. Coronin 1B coordinates Arp2/3 complex and cofilin activities at the leading edge. Cell (0092-8674) 2007 imex curation Cai L., Marshall TW., Uetrecht AC., Schafer DA., Bear JE. jbear@email.unc.edu 26-APR-2007: Contacted by JYOTI. orchard Only protein-protein interactions 7 human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Cai L., Marshall TW., Uetrecht AC., Schafer DA., Bear JE. jbear@email.unc.edu Accepted 2007-MAY-5 by ORCHARD. 26-APR-2007: Contacted by JYOTI. Cell (0092-8674) 2007 orchard Only protein-protein interactions 7 imex curation Coronin 1B coordinates Arp2/3 complex and cofilin activities at the leading edge. Coronin 1B coordinates Arp2/3 complex and cofilin activities at the leading edge. Cell (0092-8674) 2007 imex curation Cai L., Marshall TW., Uetrecht AC., Schafer DA., Bear JE. jbear@email.unc.edu 26-APR-2007: Contacted by JYOTI. orchard Only protein-protein interactions 7 in vitro In vitro phosphatase assay phosphatase assay predetermined predetermined participant predetermined Cai L., Marshall TW., Uetrecht AC., Schafer DA., Bear JE. jbear@email.unc.edu Accepted 2007-MAY-5 by ORCHARD. 26-APR-2007: Contacted by JYOTI. Cell (0092-8674) 2007 orchard Only protein-protein interactions 7 imex curation cor1b_rat Coronin-1B Coro1b Coronin-2 protein protein rat Rattus norvegicus (Rat) Rat MSFRKVVRQSKFRHVFGQPVKNDQCYEDIRVSRVTWDSTFCAVNPKFLAVIVEASGGGAFMVLPLNKTGRIDKAYPTVCGHTGPVLDIDWCPHNDEVIASGSEDCTVMVWQIPENGLTSPLTEPVVVLEGHTKRVGIITWHPTARNVLLSAGCDNVVLIWNVGTAEELYRLDSLHPDLIYNVSWNHNGSLFCTACKDKSVRIIDPRRGTLVAEREKAHEGARPMRAIFLADGKVFTAGFSRMSERQLALWDPENFEEPMALQELDSSNGALLPFYDPDTSVVYVCGKGDSSIRYFEITDEPPYIHFLNTFTSKEPQRGMGSMPKRGLEVSKCEIARFYKLHERKCEPIVMTVPRKSDLFQDDLYPDTAGPDAALEAEDWVSGQDADPILISLREAYVPSKQRDLKVSRRNVLSDSKPAGYSRSGVSTATAITDIPSGNLAGSGEAGKLEEVMHGLRALRVLVKEQGERISRLEEHLGRMENGDT 746EDF119680EBBF cor1b_human Coronin-1B CORO1B Coronin-2 protein protein human Homo sapiens Human MSFRKVVRQSKFRHVFGQPVKNDQCYEDIRVSRVTWDSTFCAVNPKFLAVIVEASGGGAFLVLPLSKTGRIDKAYPTVCGHTGPVLDIDWCPHNDEVIASGSEDCTVMVWQIPENGLTSPLTEPVVVLEGHTKRVGIIAWHPTARNVLLSAGCDNVVLIWNVGTAEELYRLDSLHPDLIYNVSWNHNGSLFCSACKDKSVRIIDPRRGTLVAEREKAHEGARPMRAIFLADGKVFTTGFSRMSERQLALWDPENLEEPMALQELDSSNGALLPFYDPDTSVVYVCGKGDSSIRYFEITEEPPYIHFLNTFTSKEPQRGMGSMPKRGLEVSKCEIARFYKLHERKCEPIVMTVPRKSDLFQDDLYPDTAGPEAALEAEEWVSGRDADPILISLREAYVPSKQRDLKISRRNVLSDSRPAMAPGSSHLGAPASTTTAADATPSGSLARAGEAGKLEEVMQELRALRALVKEQGDRICRLEEQLGRMENGDA DIP protein Q9BR76 original sequence version: 132 A6012FDA683ECB59 ssh1_human Protein phosphatase Slingshot homolog 1 SSH1 KIAA1298 SSH1L SSH-like protein 1 protein protein human Homo sapiens Human MALVTLQRSPTPSAASSSASNSELEAGSEEDRKLNLSLSESFFMVKGAALFLQQGSSPQGQRSLQHPHKHAGDLPQHLQVMINLLRCEDRIKLAVRLESAWADRVRYMVVVYSSGRQDTEENILLGVDFSSKESKSCTIGMVLRLWSDTKIHLDGDGGFSVSTAGRMHIFKPVSVQAMWSALQVLHKACEVARRHNYFPGGVALIWATYYESCISSEQSCINEWNAMQDLESTRPDSPALFVDKPTEGERTERLIKAKLRSIMMSQDLENVTSKEIRNELEKQMNCNLKELKEFIDNEMLLILGQMDKPSLIFDHLYLGSEWNASNLEELQGSGVDYILNVTREIDNFFPGLFAYHNIRVYDEETTDLLAHWNEAYHFINKAKRNHSKCLVHCKMGVSRSASTVIAYAMKEFGWPLEKAYNYVKQKRSITRPNAGFMRQLSEYEGILDASKQRHNKLWRQQTDSSLQQPVDDPAGPGDFLPETPDGTPESQLPFLDDAAQPGLGPPLPCCFRRLSDPLLPSPEDETGSLVHLEDPEREALLEEAAPPAEVHRPARQPQQGSGLCEKDVKKKLEFGSPKGRSGSLLQVEETEREEGLGAGRWGQLPTQLDQNLLNSENLNNNSKRSCPNGMEDDAIFGILNKVKPSYKSCADCMYPTASGAPEASRERCEDPNAPAICTQPAFLPHITSSPVAHLASRSRVPEKPASGPTEPPPFLPPAGSRRADTSGPGAGAALEPPASLLEPSRETPKVLPKSLLLKNSHCDKNPPSTEVVIKEESSPKKDMKPAKDLRLLFSNESEKPTTNSYLMQHQESIIQLQKAGLVRKHTKELERLKSVPADPAPPSRDGPASRLEASIPEESQDPAALHELGPLVMPSQAGSDEKSEAAPASLEGGSLKSPPPFFYRLDHTSSFSKDFLKTICYTPTSSSMSSNLTRSSSSDSIHSVRGKPGLVKQRTQEIETRLRLAGLTVSSPLKRSHSLAKLGSLTFSTEDLSSEADPSTVADSQDTTLSESSFLHEPQGTPRDPAATSKPSGKPAPENLKSPSWMSKS 0600B9690F0E6F9C arpc2_human Actin-related protein 2/3 complex subunit 2 ARPC2 ARC34 PRO2446 Arp2/3 complex 34 kDa subunit protein protein human Homo sapiens Human MILLEVNNRIIEETLALKFENAAAGNKPEAVEVTFADFDGVLYHISNPNGDKTKVMVSISLKFYKELQAHGADELLKRVYGSFLVNPESGYNVSLLYDLENLPASKDSIVHQAGMLKRNCFASVFEKYFQFQEEGKEGENRAVIHYRDDETMYVESKKDRVTVVFSTVFKDDDDVVIGKVFMQEFKEGRRASHTAPQVLFSHREPPLELKDTDAAVGDNIGYITFVLFPRHTNASARDNTINLIHTFRDYLHYHIKCSKAYIHTRMRAKTSDFLKVLNRARPDAEKKEMKTITGKTFSSR DIP protein O15144 original sequence version: 99 DIP protein O15144 original sequence version: 122 DIP protein O15144 original sequence version: 119 DIP protein O15144 original sequence version: 140 3BA57121BE9A05F2 arpc2_rat Actin-related protein 2/3 complex subunit 2 Arpc2 Arp2/3 complex 34 kDa subunit protein protein rat Rattus norvegicus (Rat) Rat MILLEVNNRIIEETLALKFENAAAGNKPEAVEVTFADFDGVLYHISNPNGDKTKVMVSISLKFYKELQAHGADELLKRVYGSFLVNPESGYNVSLLYDLENLPASKDSIVHQAGMLKRNCFASVFEKYFQFQEEGKEGENRAVIHYRDDETMYVESKKDRVTVVFSTVFKDDDDVVIGKVFMQEFKEGRRASHTAPQVLFSHREPPLELKDTDAAVGDNIGYITFVLFPRHTNATARDNTINLIHTFRDYLHYHIKCSKAYIHTRMRAKTSDFLKVLNRARPDAEKKEMKTITRKTFSST 3E016531BA8E0982 ssh1-coro1b 6 8 enzyme target enzyme target substrate substrate unspecified role unspecified role region opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 enzyme enzyme unspecified role unspecified role region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. dephosphorylation dephosphorylation reaction dephosphorylation Recombinant Coronin 1 B was expressed in Drosophila S2 cells or HEK-293 cells. 4 C Recombinant Coronin 1B phosphorylated in vitro with purified PKCa was efficiently dephosphorylated by SSH1L Coronin 1 B was dephosphorylated by Slingshot (SSH1L). coro1b-arpc2 4 10 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association 4 H SSH1L-myc interacted with endogenous Coronin 1B using reciprocal coimmunoprecipitations (Figure 4H). Arp2/3 complex (as reported by the p34 subunit) was detected in both the Coronin 1B and SSH1L immunoprecipitates. ssh1-arpc2 5 9 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey association association 4 H SSH1L-myc interacted with endogenous Coronin 1B using reciprocal coimmunoprecipitations (Figure 4H). Arp2/3 complex (as reported by the p34 subunit) was detected in both the Coronin 1B and SSH1L immunoprecipitates. coro1b-arpc2-1 3 7 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role prey prey association association 4 I In the first step, anti-Coronin 1B (but not control IgG) immunoprecipitated both SSH1L-GFP and Arp2/3 complex. ssh1-arpc2-1 1 11 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association 4 I SSH1L-GFP coimmunoprecipitated both Coronin 1B and Arp2/3 complex from a mock depleted lysate These interactions did not occur after Coronin 1B was immunodepleted from the cell lysate. The evidence suggests Coronin 1B, SSH1L, and Arp2/3 (as reported by the p34 subunit) exist in a ternary complex, bridged by Coronin 1B. arpc2-coro1b 3 9 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey association association 4 J We conclude that SSH1L, Coronin 1B and Arp2/3 exist in a ternary complex that is bridged by Coronin 1B. These interactions do not occur in Coronin 1B-depleted cells. The evidence suggests SSH1L, Coronin 1B and Arp2/3 (as reported by p34Arc) exist in a ternary complex, bridged by Coronin 1B. coro1b-arpc2-2 2 8 unspecified role unspecified role bait bait 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 10 unspecified role unspecified role prey prey human-leukocyte Homo sapiens leukocyte Leukocyte (white blood cell) physical association physical association 3 C purified Arp2/3 complex bound directly to wild-type Coronin 1B, but did not bind to the phosphomimetic S2D Coronin 1B mutant The Arp2/3 complex is reported by p34Arc. Recombinant Coronin 1 B is expressed in Drosphila S2 cells or HEK293 cells.