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/ The V0 sector of the V-ATPase, synaptobrevin, and synaptophysin are associated on synaptic vesicles in a Triton X-100-resistant, freeze-thawing sensitive, complex. The V0 sector of the V-ATPase, synaptobrevin, and synaptophysin are associated on synaptic vesicles in a Triton X-100-resistant, freeze-thawing sensitive, complex. J. Biol. Chem. (0021-9258) 1996 imex curation Galli T., McPherson PS., De Camilli P. Synapse - Interactions of proteins with an established role in the presynapse. Rat-brain synaptosomes were prepared using homogenisation followed by sequential ultracentrifugation. The supernatant was used following an overnight freeze thawing cycle followed by the addition of Triton X-100 prior to immunoprecipitation. tgalli@curie.fr,pietro_decamilli@quickmail.yale.edu 16-Dec-2008: Contacted by JYOTI. orchard Only protein-protein interactions rat-brain Rattus norvegicus brain brain Brain [cerebrum] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Galli T., McPherson PS., De Camilli P. Synapse - Interactions of proteins with an established role in the presynapse. Rat-brain synaptosomes were prepared using homogenisation followed by sequential ultracentrifugation. The supernatant was used following an overnight freeze thawing cycle followed by the addition of Triton X-100 prior to immunoprecipitation. tgalli@curie.fr,pietro_decamilli@quickmail.yale.edu Accepted 2008-NOV-27 by ORCHARD. 16-Dec-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 1996 orchard Only protein-protein interactions imex curation The V0 sector of the V-ATPase, synaptobrevin, and synaptophysin are associated on synaptic vesicles in a Triton X-100-resistant, freeze-thawing sensitive, complex. The V0 sector of the V-ATPase, synaptobrevin, and synaptophysin are associated on synaptic vesicles in a Triton X-100-resistant, freeze-thawing sensitive, complex. J. Biol. Chem. (0021-9258) 1996 imex curation Galli T., McPherson PS., De Camilli P. Synapse - Interactions of proteins with an established role in the presynapse. Rat-brain synaptosomes were prepared using homogenisation followed by sequential ultracentrifugation. The supernatant was used following an overnight freeze thawing cycle followed by the addition of Triton X-100 prior to immunoprecipitation. tgalli@curie.fr,pietro_decamilli@quickmail.yale.edu 16-Dec-2008: Contacted by JYOTI. orchard Only protein-protein interactions rat-pc12 Rattus norvegicus PC12 cells pc12 Rat adrenal pheochromocytoma cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Galli T., McPherson PS., De Camilli P. Synapse - Interactions of proteins with an established role in the presynapse. Rat-brain synaptosomes were prepared using homogenisation followed by sequential ultracentrifugation. The supernatant was used following an overnight freeze thawing cycle followed by the addition of Triton X-100 prior to immunoprecipitation. tgalli@curie.fr,pietro_decamilli@quickmail.yale.edu Accepted 2008-NOV-27 by ORCHARD. 16-Dec-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 1996 orchard Only protein-protein interactions imex curation The V0 sector of the V-ATPase, synaptobrevin, and synaptophysin are associated on synaptic vesicles in a Triton X-100-resistant, freeze-thawing sensitive, complex. The V0 sector of the V-ATPase, synaptobrevin, and synaptophysin are associated on synaptic vesicles in a Triton X-100-resistant, freeze-thawing sensitive, complex. J. Biol. Chem. (0021-9258) 1996 imex curation Galli T., McPherson PS., De Camilli P. Synapse - Interactions of proteins with an established role in the presynapse. Rat-brain synaptosomes were prepared using homogenisation followed by sequential ultracentrifugation. The supernatant was used following an overnight freeze thawing cycle followed by the addition of Triton X-100 prior to immunoprecipitation. tgalli@curie.fr,pietro_decamilli@quickmail.yale.edu 16-Dec-2008: Contacted by JYOTI. orchard Only protein-protein interactions rat-brain Rattus norvegicus brain brain Brain [cerebrum] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Galli T., McPherson PS., De Camilli P. Synapse - Interactions of proteins with an established role in the presynapse. Rat-brain synaptosomes were prepared using homogenisation followed by sequential ultracentrifugation. The supernatant was used following an overnight freeze thawing cycle followed by the addition of Triton X-100 prior to immunoprecipitation. tgalli@curie.fr,pietro_decamilli@quickmail.yale.edu Accepted 2008-NOV-27 by ORCHARD. 16-Dec-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 1996 orchard Only protein-protein interactions imex curation stx1a_rat Syntaxin-1A Synaptotagmin-associated 35 kDa protein Neuron-specific antigen HPC-1 Stx1a Sap protein protein rat Rattus norvegicus (Rat) Rat MKDRTQELRTAKDSDDDDDVTVTVDRDRFMDEFFEQVEEIRGFIDKIAENVEEVKRKHSAILASPNPDEKTKEELEELMSDIKKTANKVRSKLKSIEQSIEQEEGLNRSSADLRIRKTQHSTLSRKFVEVMSEYNATQSDYRERCKGRIQRQLEITGRTTTSEELEDMLESGNPAIFASGIIMDSSISKQALSEIETRHSEIIKLENSIRELHDMFMDMAMLVESQGEMIDRIEYNVEHAVDYVERAVSDTKKAVKYQSKARRKKIMIIICCVILGIIIASTIGGIFG DIP protein P32851 original sequence version: 74 DIP protein P32851 original sequence version: 78 DIP protein P32851 original sequence version: 142 DIP protein P32851 original sequence version: 138 DIP protein P32851 original sequence version: 89 DIP protein P32851 original sequence version: 115 DIP protein P32851 original sequence version: 134 DIP protein P32851 original sequence version: 121 DIP protein P32851 original sequence version: 123 DIP protein P32851 original sequence version: 150 DIP protein P32851 original sequence version: 156 DIP protein P32851 original sequence version: 163 DIP protein P32851 original sequence version: 126 DIP protein P32851 original sequence version: 124 DIP protein P32851 original sequence version: 158 8F929F489E323CAA vpp1_rat V-type proton ATPase 116 kDa subunit a1 Atp6v0a1 Atp6n1 Vpp1 Vacuolar proton translocating ATPase 116 kDa subunit a isoform 1 Clathrin-coated vesicle/synaptic vesicle proton pump 116 kDa subunit Vacuolar proton pump subunit 1 Vacuolar adenosine triphosphatase subunit Ac116 protein protein rat Rattus norvegicus (Rat) Rat MGELFRSEEMTLAQLFLQSEAAYCCVSELEELGKVQFRDLNPDVNVFQRKFVNEVRRCEEMDRKLRFVEKEIRKANIPIMDTGENPEVPFPRDMIDLEANFEKIENELKEINTNQEALKRNFLELTELKFILRKTQQFFDEMADPDLLEESSSLLEPNEMGRGAPLRLGFVAGVINRERIPTFERMLWRVCRGNVFLRQAEIENPLEDPVTGDYVHKSVFIIFFQGDQLKNRVKKICEGFRASLYPCPETPQERKEMASGVNTRIDDLQMVLNQTEDHRQRVLQAAAKNIRVWFIKVRKMKAIYHTLNLCNIDVTQKCLIAEVWCPVTDLDSIQFALRRGTEHSGSTVPSILNRMQTNQTPPTYNKTNKFTHGFQNIVDAYGIGTYREINPAPYTVITFPFLFAVMFGDFGHGILMTLFAVWMVLRESRILSQKNENEMFSMVFSGRYIILLMGLFSIYTGLIYNDCFSKSLNIFGSSWSVRPMFTIGNWTEETLLGSSVLQLNPAIPGVFGGPYPFGIDPIWNIATNKLTFLNSFKMKMSVILGIIHMLFGVSLSLFNHIYFKKPLNIYFGFIPEIIFMSSLFGYLVILIFYKWTAYDAHSSRNAPSLLIHFINMFLFSYPESGNAMLYSGQKGIQCFLIVVAMLCVPWMLLFKPLILRHQYLRKKHLGTLNFGGIRVGNGPTEEDAEIIQHDQLSTHSEDAEEPTEDEVFDFGDTMVHQAIHTIEYCLGCISNTASYLRLWALSLAHAQLSEVLWTMVIHIGLHVRSLAGGLGLFFIFAAFATLTVAILLIMEGLSAFLHALRLHWVEFQNKFYTGTGFKFLPFSFEHIREGKFDE D25FAD918638D7EE syph_rat Synaptophysin Major synaptic vesicle protein p38 Syp protein protein rat Rattus norvegicus (Rat) Rat MDVVNQLVAGGQFRVVKEPLGFVKVLQWVFAIFAFATCGSYTGELRLSVECANKTESALNIEVEFEYPFRLHQVYFDAPSCVKGGTTKIFLVGDYSSSAEFFVTVAVFAFLYSMGALATYIFLQNKYRENNKGPMMDFLATAVFAFMWLVSSSAWAKGLSDVKMATDPENIIKEMPMCRQTGNTCKELRDPVTSGLNTSVVFGFLNLVLWVGNLWFVFKETGWAAPFMRAPPGAPEKQPAPGDAYGDAGYGQGPGGYGPQDSYGPQGGYQPDYGQPASGGGGYGPQGDYGQQGYGQQGAPTSFSNQM A245E4F70BF0EAF0 syt1_rat Synaptotagmin-1 Syt1 Synaptotagmin I p65 protein protein rat Rattus norvegicus (Rat) Rat MVSASHPEALAAPVTTVATLVPHNATEPASPGEGKEDAFSKLKQKFMNELHKIPLPPWALIAIAIVAVLLVVTCCFCVCKKCLFKKKNKKKGKEKGGKNAINMKDVKDLGKTMKDQALKDDDAETGLTDGEEKEEPKEEEKLGKLQYSLDYDFQNNQLLVGIIQAAELPALDMGGTSDPYVKVFLLPDKKKKFETKVHRKTLNPVFNEQFTFKVPYSELGGKTLVMAVYDFDRFSKHDIIGEFKVPMNTVDFGHVTEEWRDLQSAEKEEQEKLGDICFSLRYVPTAGKLTVVILEAKNLKKMDVGGLSDPYVKIHLMQNGKRLKKKKTTIKKNTLNPYYNESFSFEVPFEQIQKVQVVVTVLDYDKIGKNDAIGKVFVGYNSTGAELRHWSDMLANPRRPIAQWHTLQVEEEVDAMLAVKK DIP protein P21707 original sequence version: 71 DIP protein P21707 original sequence version: 76 DIP protein P21707 original sequence version: 141 DIP protein P21707 original sequence version: 112 DIP protein P21707 original sequence version: 145 DIP protein P21707 original sequence version: 160 06CE28F04C97A722 vamp2_rat Vesicle-associated membrane protein 2 Vamp2 Syb2 Synaptobrevin-2 protein protein rat Rattus norvegicus (Rat) Rat MSATAATVPPAAPAGEGGPPAPPPNLTSNRRLQQTQAQVDEVVDIMRVNVDKVLERDQKLSELDDRADALQAGASQFETSAAKLKRKYWWKNLKMMIILGVICAIILIIIIVYFST DIP protein P63045 original sequence version: 95 DIP protein P63045 original sequence version: 90 DIP protein P63045 original sequence version: 64 DIP protein P63045 original sequence version: 62 DIP protein P63045 original sequence version: 85 DIP protein P63045 original sequence version: 104 DIP protein P63045 original sequence version: 77 DIP protein P63045 original sequence version: 75 DIP protein P63045 original sequence version: 106 4A0D0D56B5409D0A snp25_rat Synaptosomal-associated protein 25 Snap25 Snap Synaptosomal-associated 25 kDa protein Super protein protein protein rat Rattus norvegicus (Rat) Rat MAEDADMRNELEEMQRRADQLADESLESTRRMLQLVEESKDAGIRTLVMLDEQGEQLERIEEGMDQINKDMKEAEKNLTDLGKFCGLCVCPCNKLKSSDAYKKAWGNNQDGVVASQPARVVDEREQMAISGGFIRRVTNDARENEMDENLEQVSGIIGNLRHMALDMGNEIDTQNRQIDRIMEKADSNKTRIDEANQRATKMLGSG DIP protein P60881 original sequence version: 105 DIP protein P60881 original sequence version: 40 DIP protein P60881 original sequence version: 106 DIP protein P60881 original sequence version: 78 DIP protein P60881 original sequence version: 75 DIP protein P60881 original sequence version: 100 DIP protein P60881 original sequence version: 120 DIP protein P60881 original sequence version: 131 DIP protein P60881 original sequence version: 93 FBED2B082A4CB6A6 vatl_rat V-type proton ATPase 16 kDa proteolipid subunit Atp6v0c Atp6c Atp6l Atpl Vacuolar proton pump 16 kDa proteolipid subunit protein protein rat Rattus norvegicus (Rat) Rat MADIKNNPEYSSFFGVMGASSAMVFSAMGAAYGTAKSGTGIAAMSVMRPELIMKSIIPVVMAGIIAIYGLVVAVLIANSLTDGITLYRSFLQLGAGLSVGLSGLAAGFAIGIVGDAGVRGTAQQPRLFVGMILILIFAEVLGLYGLIVALILSTK 880C280C5AEB0C5C q5m7t6_rat V-type proton ATPase subunit Atp6v0d1 rCG_51062 protein protein rat Rattus norvegicus (Rat) Rat MSFFPELYFNVDNGYLEGLVRGLKAGVLSQADYLNLVQCETLEDLKLHLQSTDYGNFLANEASPLTVSVIDDKLKEKMVVEFRHMRNHAYEPLASFLDFITYSYMIDNVILLITGTLHQRSIAELVPKCHPLGSFEQMEAVNIAQTPAELYNAILVDTPLAAFFQDCISEQDLDEMNIEIIRNTLYKAYLESFYKFCTLLGGTTADAMCPILEFEADRRAFIITINSFGTELSKEDRAKLFPHCGRLYPEGLAQLARADDYEQVKNVADYYPEYKLLFEGAGSNPGDKTLEDRFFEHEVKLNKLAFLNQFHFGVFYAFVKLKEQECRNIVWIAECIAQRHRAKIDNYIPIF 62CDF67B982124C9 vamp2-stxa1 1 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 4 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association 3C, 2D When frozen-thawed extract was used, the major proteins which coprecipitated with synaptobrevin 2 were SNAP-25 and syntaxin 1 (Fig. 1, a and b, and Fig. 3), in addition to a band of approximately 85 kDa vamp2-syp-1 3 11 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 4 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association 1A, 3 when freshly made Triton X-100 extract was used, synaptophysin (Fig. 1, a and b, see also Fig. 3) and a protein migrating at approximately 80 kDa were the major bands visible by protein staining in the immunoprecipitates. In addition, a 10-kDa protein (p10) and several other minor bands of higher molecular mass were visible. stxa1-vamp2 1 4 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association 4 freeze-thawing treatment which induces dissociation of synaptobrevin 2 from synaptophysin and V-ATPase also induces dissociation of synaptophysin from V-ATPase. stxa1-vamp2-1 3 4 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association 4 vamp2-syp-2 3 10 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association 4 We confirmed the specificity of the association between synaptobrevin 2, synaptophysin, and the V-ATPase by performing immunoprecipitation experiments using monoclonal antibodies against synaptophysin (Cl 7.2) and syntaxin 1 (HPC-1) (Fig. 4). vamp2-syp-3 2 8 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey physical association physical association 5 The c subunit of the V-ATPase was found to be associated with synaptobrevin 2 and synaptophysin also in freshly prepared Triton X-100 extracts of PC12 cells. vamp2-syp-4 2 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey physical association physical association 5 The c subunit of the V-ATPase was found to be associated with synaptobrevin 2 and synaptophysin also in freshly prepared Triton X-100 extracts of PC12 cells.