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/ Characterization of a multidomain adaptor protein, p140Cap, as part of a pre-synaptic complex. Characterization of a multidomain adaptor protein, p140Cap, as part of a pre-synaptic complex. J. Neurochem. (0022-3042) 2008 imex curation Ito H., Atsuzawa K., Sudo K., Di Stefano P., Iwamoto I., Morishita R., Takei S., Semba R., Defilippi P., Asano T., Usuda N., Nagata K. Synapse - Interactions of proteins with an established role in the presynapse. knagata@inst-hsc.jp 13-Sept-2010: Contacted by JYOTI. Human brain cDNA. jyoti Only protein-protein interactions chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Ito H., Atsuzawa K., Sudo K., Di Stefano P., Iwamoto I., Morishita R., Takei S., Semba R., Defilippi P., Asano T., Usuda N., Nagata K. Synapse - Interactions of proteins with an established role in the presynapse. knagata@inst-hsc.jp Accepted 2010-JUL-30 by JYOTI. 13-Sept-2010: Contacted by JYOTI. Human brain cDNA. J. Neurochem. (0022-3042) 2008 jyoti Only protein-protein interactions imex curation Characterization of a multidomain adaptor protein, p140Cap, as part of a pre-synaptic complex. Characterization of a multidomain adaptor protein, p140Cap, as part of a pre-synaptic complex. J. Neurochem. (0022-3042) 2008 imex curation Ito H., Atsuzawa K., Sudo K., Di Stefano P., Iwamoto I., Morishita R., Takei S., Semba R., Defilippi P., Asano T., Usuda N., Nagata K. Synapse - Interactions of proteins with an established role in the presynapse. knagata@inst-hsc.jp 13-Sept-2010: Contacted by JYOTI. Human brain cDNA. jyoti Only protein-protein interactions chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 fluorescence imaging fluorescence microscopy fluorescence imaging anti tag immunost anti tag immunostaining anti tag immunost Ito H., Atsuzawa K., Sudo K., Di Stefano P., Iwamoto I., Morishita R., Takei S., Semba R., Defilippi P., Asano T., Usuda N., Nagata K. Synapse - Interactions of proteins with an established role in the presynapse. knagata@inst-hsc.jp Accepted 2010-JUL-30 by JYOTI. 13-Sept-2010: Contacted by JYOTI. Human brain cDNA. J. Neurochem. (0022-3042) 2008 jyoti Only protein-protein interactions imex curation Characterization of a multidomain adaptor protein, p140Cap, as part of a pre-synaptic complex. Characterization of a multidomain adaptor protein, p140Cap, as part of a pre-synaptic complex. J. Neurochem. (0022-3042) 2008 imex curation Ito H., Atsuzawa K., Sudo K., Di Stefano P., Iwamoto I., Morishita R., Takei S., Semba R., Defilippi P., Asano T., Usuda N., Nagata K. Synapse - Interactions of proteins with an established role in the presynapse. knagata@inst-hsc.jp 13-Sept-2010: Contacted by JYOTI. Human brain cDNA. jyoti Only protein-protein interactions rat-brain-cerebellum Rattus norvegicus brain_cerebellum [Cerebellar] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Ito H., Atsuzawa K., Sudo K., Di Stefano P., Iwamoto I., Morishita R., Takei S., Semba R., Defilippi P., Asano T., Usuda N., Nagata K. Synapse - Interactions of proteins with an established role in the presynapse. knagata@inst-hsc.jp Accepted 2010-JUL-30 by JYOTI. 13-Sept-2010: Contacted by JYOTI. Human brain cDNA. J. Neurochem. (0022-3042) 2008 jyoti Only protein-protein interactions imex curation Characterization of a multidomain adaptor protein, p140Cap, as part of a pre-synaptic complex. Characterization of a multidomain adaptor protein, p140Cap, as part of a pre-synaptic complex. J. Neurochem. (0022-3042) 2008 imex curation Ito H., Atsuzawa K., Sudo K., Di Stefano P., Iwamoto I., Morishita R., Takei S., Semba R., Defilippi P., Asano T., Usuda N., Nagata K. Synapse - Interactions of proteins with an established role in the presynapse. knagata@inst-hsc.jp 13-Sept-2010: Contacted by JYOTI. Human brain cDNA. jyoti Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Ito H., Atsuzawa K., Sudo K., Di Stefano P., Iwamoto I., Morishita R., Takei S., Semba R., Defilippi P., Asano T., Usuda N., Nagata K. Synapse - Interactions of proteins with an established role in the presynapse. knagata@inst-hsc.jp Accepted 2010-JUL-30 by JYOTI. Human brain cDNA. J. Neurochem. (0022-3042) 2008 jyoti Only protein-protein interactions imex curation syph_rat Synaptophysin Major synaptic vesicle protein p38 Syp protein protein rat Rattus norvegicus (Rat) Rat MDVVNQLVAGGQFRVVKEPLGFVKVLQWVFAIFAFATCGSYTGELRLSVECANKTESALNIEVEFEYPFRLHQVYFDAPSCVKGGTTKIFLVGDYSSSAEFFVTVAVFAFLYSMGALATYIFLQNKYRENNKGPMMDFLATAVFAFMWLVSSSAWAKGLSDVKMATDPENIIKEMPMCRQTGNTCKELRDPVTSGLNTSVVFGFLNLVLWVGNLWFVFKETGWAAPFMRAPPGAPEKQPAPGDAYGDAGYGQGPGGYGPQDSYGPQGGYQPDYGQPASGGGGYGPQGDYGQQGYGQQGAPTSFSNQM A245E4F70BF0EAF0 wasl_human Neural Wiskott-Aldrich syndrome protein WASL protein protein human Homo sapiens Human MSSVQQQPPPPRRVTNVGSLLLTPQENESLFTFLGKKCVTMSSAVVQLYAADRNCMWSKKCSGVACLVKDNPQRSYFLRIFDIKDGKLLWEQELYNNFVYNSPRGYFHTFAGDTCQVALNFANEEEAKKFRKAVTDLLGRRQRKSEKRRDPPNGPNLPMATVDIKNPEITTNRFYGPQVNNISHTKEKKKGKAKKKRLTKADIGTPSNFQHIGHVGWDPNTGFDLNNLDPELKNLFDMCGISEAQLKDRETSKVIYDFIEKTGGVEAVKNELRRQAPPPPPPSRGGPPPPPPPPHNSGPPPPPARGRGAPPPPPSRAPTAAPPPPPPSRPSVAVPPPPPNRMYPPPPPALPSSAPSGPPPPPPSVLGVGPVAPPPPPPPPPPPGPPPPPGLPSDGDHQVPTTAGNKAALLDQIREGAQLKKVEQNSRPVSCSGRDALLDQIRQGIQLKSVADGQESTPPTPAPTSGIVGALMEVMQKRSKAIHSSDEDEDEDDEEDFEDDDEWED DIP protein O00401 original sequence version: 55 DIP protein O00401 original sequence version: 147 DIP protein O00401 original sequence version: 150 DIP protein O00401 original sequence version: 136 DIP protein O00401 original sequence version: 138 DIP protein O00401 original sequence version: 125 DIP protein O00401 original sequence version: 112 DIP protein O00401 original sequence version: 126 B5895C6055E23049 srcn1_rat SRC kinase signaling inhibitor 1 Srcin1 Snip p140Cap SNAP-25-interacting protein P140 p130Cas-associated protein protein protein rat Rattus norvegicus (Rat) Rat MQPWQCLRRFALAWWERTAEGRARSPREEVGPRDPGGRGEPDPERSSPPMLSADDAEYPREYRTLGGGGSGGSGGRRFSNVGLVHTSERRHTVIAAQSLEALSGLQKADADRKRDAFMDHLKSKYPQHALALRGQQDRMREQQPNYWSFKTRSSRHTQGAQPGLADQAAKLSYASAESLETMSEAELPLGFSRMNRFRQSLPLSRSASQTKLRSPGVLFLQFGEETRRVHITHEVSSLDTLHALIAHMFPQKLTMGMLKSPNTAILIKDEARNVFYELEDVRDIQDRSIIKIYRKEPLYAAFPGSHLTNGDLRREMVYASRESSPTRRLNNLSPASHLASSSPPPGLPSGLPSGLPSGSPSRSRLSYAGGRPPSYAGSPVHHAAERLGGAPTSQGVSPSPSAILERRDVKPDEDLAGKAGGMVLVKGEGLYADPYGLLHEGRLSLAAAAGDPFAYPGAGGLYKRGSVRSLSTYSAAALQSDLEDSLYKAGAGGPLYGDGYGFRLPPSSPQKLADVSAPSGGPPPPHSPYSGPPSRGSPVRQSFRKDSGSSSVFAESPGGKARSTGASTAGAPPSELFPGPGERSLVGFGPPVPAKDTETRERMEAMEKQIASLTGLVQSALLRGSEPETPSEKIEGSNGAATPSAPVCGSGSRSSGATPVSGPPPPAVSSTPAGQPTAVSRLQMQMHLRGLQNSASDLRGQLQQLRKLQLQNQESVRALLKRTEAELSMRVSEAARRQEDPLQRQRTLVEEERLRYLNDEELITQQLNDLEKSVEKIQRDVAHNHRLVPGPELEEKALVLKQLGETLTELKAHFPGLQSKMRVVLRVEVEAVKFLKEEPQRLDGLLKRCRVVTDTLAQIRRQVDEGVWPPPNNLLNQSPKKVAAETDFSKGLDFEIPPPSPPLNLHELSGPAEGTPLTPKSGNPTKGLDAPSKRNMDKAVSVEAAERDWEEKRAALTQYSAKDINRLLEETQAELLKAIPDLDCASKTHPGPTPTPDHKPPKAPHGQKAAPRTEPSGRRGSDELTVPRYRTEKPSKSPPPPPPRRSFPSSHGLTTTRTGEVVVTSKKDSVFIKKAESEELEIQKPQVKLRRAVSEVVRPASTPPIMASAIKDEDDEERIIAELESGGGSVPPMKVVTPGASRLKAAQGPAGSPDKGKHGKQRTEYMRIQAQQQVRVGYQAPRPLEGCTPRLCYMPLS 8C9A2E630FB31C33 q8cj75_rat protein protein rat Rattus norvegicus (Rat) Rat MQGLPAGLSLDDFIPGHLRAHIGASSRGTRVPVIRNGGSNTLNFQFHDPAPRTVCNGCPPPRRDGSQHPDPAWYQTWPGPGSQPSVSPKQPAPQNAQNWSATWTKDSKRRDKRWVKYEGIGPVDESGMPIAPRSSVDSPRDWYRRMFQQIHRKMPDLQLDWTLEDPPKVVSSRASSADPRHPGPQQRPASRPGTTGTSSGRNWNHSEETSRNTLNYNLGPSSRLHASNQVPRHREKVENVWTEDSWNQFLHELETGHKPKKSLVDDPVEKPAQPIEVLLERELAKLSAELDKDLRAIETRLPSPKNSQAPRRPLEQPGPEQQPSARLSSAWRPNSPHAPYFGSSRPLSPHRMADGGGSPFLGRRDFVYPSSAREPSASERGSSPARKEEKKRKAARLKFDFQAQSPKELTLQKGDIVYIHKEVDKNWLEGEHHGRLGIFPANYVEVLPADEIPKPIKP 8EFBE5D97F56E39C o60504-1 Vinexin SORBS3 SCAM1 SH3-containing adapter molecule 1 Sorbin and SH3 domain-containing protein 3 protein protein human Homo sapiens Human MQGPPRSLRAGLSLDDFIPGHLQSHIGSSSRGTRVPVIRNGGSNTLNFQFHDPAPRTVCNGGYTPRRDASQHPDPAWYQTWPGPGSKPSASTKIPASQHTQNWSATWTKDSKRRDKRWVKYEGIGPVDESGMPIAPRSSVDRPRDWYRRMFQQIHRKMPDLQLDWTFEEPPRDPRHLGAQQRPAHRPGPATSSSGRSWDHSEELPRSTFNYRPGAFSTVLQPSNQVLRRREKVDNVWTEESWNQFLQELETGQRPKKPLVDDPGEKPSQPIEVLLERELAELSAELDKDLRAIETRLPSPKSSPAPRRAPEQRPPAGPASAWSSSYPHAPYLGSARSLSPHKMADGGSPFLGRRDFVYPSSTRDPSASNGGGSPARREEKKRKAARLKFDFQAQSPKELTLQKGDIVYIHKEVDKNWLEGEHHGRLGIFPANYVEVLPADEIPKPIKPPTYQVLEYGEAVAQYTFKGDLEVELSFRKGEHICLIRKVNENWYEGRITGTGRQGIFPASYVQVSREPRLRLCDDGPQLPTSPRLTAAARSARHPSSPSALRSPADPIDLGGQTSPRRTGFSFPTQEPRPQTQNLGTPGPALSHSRGPSHPLDLGTSSPNTSQIHWTPYRAMYQYRPQNEDELELREGDRVDVMQQCDDGWFVGVSRRTQKFGTFPGNYVAPV 2D8A8718E0367BE9 o60504-2 Vinexin SORBS3 SCAM1 SH3-containing adapter molecule 1 Sorbin and SH3 domain-containing protein 3 protein protein human Homo sapiens Human MADGGSPFLGRRDFVYPSSTRDPSASNGGGSPARREEKKRKAARLKFDFQAQSPKELTLQKGDIVYIHKEVDKNWLEGEHHGRLGIFPANYVEVLPADEIPKPIKPPTYQVLEYGEAVAQYTFKGDLEVELSFRKGEHICLIRKVNENWYEGRITGTGRQGIFPASYVQVSREPRLRLCDDGPQLPTSPRLTAAARSARHPSSPSALRSPADPIDLGGQTSPRRTGFSFPTQEPRPQTQNLGTPGPALSHSRGPSHPLDLGTSSPNTSQIHWTPYRAMYQYRPQNEDELELREGDRVDVMQQCDDGWFVGVSRRTQKFGTFPGNYVAPV Contains a phosphoserine at position 6. Initiator Met-1 is removed. Contains a N-acetylalanine at position 2. 852B1FDC07859AC2 vinex_human Vinexin SCAM1 SH3-containing adapter molecule 1 Sorbin and SH3 domain-containing protein 3 SORBS3 protein protein human Homo sapiens Human MQGPPRSLRAGLSLDDFIPGHLQSHIGSSSRGTRVPVIRNGGSNTLNFQFHDPAPRTVCNGGYTPRRDASQHPDPAWYQTWPGPGSKPSASTKIPASQHTQNWSATWTKDSKRRDKRWVKYEGIGPVDESGMPIAPRSSVDRPRDWYRRMFQQIHRKMPDLQLDWTFEEPPRDPRHLGAQQRPAHRPGPATSSSGRSWDHSEELPRSTFNYRPGAFSTVLQPSNQVLRRREKVDNVWTEESWNQFLQELETGQRPKKPLVDDPGEKPSQPIEVLLERELAELSAELDKDLRAIETRLPSPKSSPAPRRAPEQRPPAGPASAWSSSYPHAPYLGSARSLSPHKMADGGSPFLGRRDFVYPSSTRDPSASNGGGSPARREEKKRKAARLKFDFQAQSPKELTLQKGDIVYIHKEVDKNWLEGEHHGRLGIFPANYVEVLPADEIPKPIKPPTYQVLEYGEAVAQYTFKGDLEVELSFRKGEHICLIRKVNENWYEGRITGTGRQGIFPASYVQVSREPRLRLCDDGPQLPTSPRLTAAARSARHPSSPSALRSPADPIDLGGQTSPRRTGFSFPTQEPRPQTQNLGTPGPALSHSRGPSHPLDLGTSSPNTSQIHWTPYRAMYQYRPQNEDELELREGDRVDVMQQCDDGWFVGVSRRTQKFGTFPGNYVAPV 2D8A8718E0367BE9 b4dvr4_human protein protein human Homo sapiens Human MGAMGGQSGLGVTRVGCGAPGWRLSLCAPCSPASAWSSSYPHAPYLGSARSLSPHKMADGGSPFLGRRDFVYPSSTRDPSASNGGGSPARREEKKRKAARLKFDFQAQSPKELTLQKGDIVYIHKEVDKNWLEGEHHGRLGIFPANYVEVLPADEIPKPIKPPTYQVLEYGEAVAQYTFKGDLEVELSFRKGEHICLIRKVNENWYEGRITGTGRQGIFPASYVQVSREPRLRLCDDGPQLPTSPRLTAAARSARHPSSPSALRSPADPTDLGGQTSPRRTGFSFPTQEPRPQTQNLGTPGPALSHSRGPSHPLDLGTSSPNTSQIHWTPYRAMYQYRPQNEDELELREGDRVDVMQQCDDGWFVGVSRRTQKFGTFPGNYVAPV A39423DB4FA9872C q9qwi6-2 SRC kinase signaling inhibitor 1 1b Srcin1 Kiaa1684 P140 SNAP-25-interacting protein p130Cas-associated protein p140Cap protein protein mouse Mus musculus Mouse MGNAPSQDPERSSPPMLSADDAEYPREYRTLGGGGGGGSGGRRFSNVGLVHTSERRHTVIAAQSLEALSGLQKADADRKRDAFMDHLKSKYPQHALALRGQQDRMREQVGGWTVDPVCLLSSLCSHLHGDSTPSGAGQPAQQPNYWSFKTRSSRHTQGAQPGLADQAAKLSYASAESLETMSEAELPLGFSRMNRFRQSLPLSRSASQTKLRSPGVLFLQFGEETRRVHITHEVSSLDTLHALIAHMFPQKLTMGMLKSPNTAILIKDEARNVFYELEDVRDIQDRSIIKIYRKEPLYAAFPGSHLTNGDLRREMVYASRESSPTRRLNNLSPASHLASSSPPPGLPSGLPSGLPSGSPSRSRLSYAGGRPPSYAGSPVHHAAERLGGAPTGQGVSPSPSAILERRDVKPDEDLAGKAGGMVLVKGEGLYADPYGLLHEGRLSLAAAAETHSHTRARAACTSGVPCALSAPTPLPRCSPTWRTRCTRRALAALYGDGYGFRLPPSSPQKLADVSAPSGGPPPPHSPYSGPPSRGSPVRQSFRKDSGSSSVFAESPGGKARSTGSASTAGAPPSELFPGPGERSLVGFGPPVPAKDTETRERMEAMEKQIASLTGLVQSALLRGSEPETPSEKVEGSNGAATPSAPVCGSGSKSSGATPVSGPPPPSASSTPAGQPTAVSRLQMQLHLRGLQNSASDLRGQLQQLRNVQLQNQESVRALLKPTEADVSMRVSEAARRQEDPLQRQRTLVEEERLRYLNDEELITQQLNDLEKSVEKIQRDVAHNHRLMPGPELEEKALVLKQLGETLTELKAHFPGLQSKMRVVLRVEVEAVKFLKEEPQRLDGLLKRCRGVTDTLAQIRRQVDEGMWPPPNNLLNQSPKKVAAETDFSKGLDFEIPPPSPPLNLHELSGPAEGTPLTPKSTNPTKCLDASSKRNTDKAVSVEAAERDWEEKRAALTQYSAKDINRLLEETQAELLKAIPDLDCASKTHPGPAPTPDHKPPKAPHGQKAAPRTEPSGRRGSDELTVPRYRTEKPSKSPPPPPPRRSFPSSHGLTTTRTGEVVVTSKKDSVFIKKAESEELEVQKPQVKLRRAVSEVVRPASTPPIMASAIKDEDDEERIIAELESGGSSVPPMKVVTPGASRLKAAQGPAGSPDKGKHGKQRTEYMRIQAQQQATKPSKEVSGPNETSSPGSEKPSGSRTSIPVLTSFGARNSSISF 67E3F46F8DB2BF60 p140-sorbs3-1 4 11 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Using p140Cap-C containing the C-terminally located Pro-rich motif as a bait (Fig. 7a), we performed yeast two-hybrid screening of a human brain cDNA library (Nagata et al. 1998) and identified vinexin, a member of a novel adaptor protein family. Fig. 7A p140-sorbs3-2 1 12 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-2658658]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 341-341, sequence length 329 [EBI-2658658]252-341 13 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 physical association physical association As shown in Fig. 7b, p140Cap was immunoprecipitated with vinexinβ. Fig. 7B p140-sorbs3-3 1 13 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 10 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Lysates from COS7 cells expressing Myc-p140Cap or -p140Cap-Pro with or without Flag-vinexinβ were immunoprecipitated with M2 and analyzed as in (b). Fig. 7C p140-sorbs3 3 7 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association When endogenous p140Cap was immunoprecipitated from adult rat brain extracts, the protein was found to form complexes with vinexinα/γ, isoforms dominantly expressed in brain tissues (Fig. 7d). Fig. 7D p140-sorbs3-4 2 9 unspecified role unspecified role neutral component neutral component region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 13 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization When the two proteins were co-expressed, vinexinα came to co-localize with p140Cap in the perinuclear region, especially in cells where vinexinα was highly expressed (Fig. 8a, bottom panels) Fig. 8A p140-sorbs3-5 1 13 competitor competitor competitor prey prey 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 bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association As shown in Fig. 8b, this N-WASP-vinexin interaction was suppressed when p140Cap was co-expressed under the conditions used. Fig. 8B p140-sorbs3-6 1 13 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 10 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figs. 8B, 9D wasl-sorbs3-1 1 10 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Neuronal (N)-WASP has been shown to bind with the third SH3 domain of vinexin Fig. 8B p140-syp 3 7 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey physical association physical association Interestingly, immunocomplexes from rat brain extracts contained a synaptic vesicle membrane protein, synaptophysin (Fig. 9a), Fig. 9A p140-syp-1 1 5 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 13 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 physical association physical association Fig. 9B We then confirmed the interaction of Myc-p140Cap with Flag-synaptophysin in COS7 cell expression experiments (Fig. 9b). The synaptophysin-p140Cap interaction was lost when synaptophysin delta-N or delta-C mutant was used instead of the wild type. p140-syp-2 1 5 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 13 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 centre region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association Fig. 9C As for p140Cap, the center region containing coiled-coil domains and a charged sequence region was found to be important to bind with synaptophysin (Fig. 9c). p140-sorbs3-7 1 5 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 10 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 13 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 physical association physical association Fig. 9D Indeed, p140Cap, synaptophysin, and vinexin formed a ternary complex in COS7 cells (Fig. 9d).