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/ Sequential tethering of Golgins and catalysis of SNAREpin assembly by the vesicle-tethering protein p115. Sequential tethering of Golgins and catalysis of SNAREpin assembly by the vesicle-tethering protein p115. J. Cell Biol. (0021-9525) 2002 imex curation Shorter J., Beard M.B., Seemann J., Dirac-Svejstrup A.B., Warren G. graham.warren@yale.edu 22-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions rat-liver rattus norvegicus liver Rat liver liver anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Shorter J., Beard M.B., Seemann J., Dirac-Svejstrup A.B., Warren G. graham.warren@yale.edu Accepted 2015-JAN-05 by ORCHARD J. Cell Biol. (0021-9525) 2002 Only protein-protein interactions imex curation Sequential tethering of Golgins and catalysis of SNAREpin assembly by the vesicle-tethering protein p115. Sequential tethering of Golgins and catalysis of SNAREpin assembly by the vesicle-tethering protein p115. J. Cell Biol. (0021-9525) 2002 imex curation Shorter J., Beard M.B., Seemann J., Dirac-Svejstrup A.B., Warren G. graham.warren@yale.edu 22-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Shorter J., Beard M.B., Seemann J., Dirac-Svejstrup A.B., Warren G. graham.warren@yale.edu Accepted 2015-JAN-05 by ORCHARD J. Cell Biol. (0021-9525) 2002 Only protein-protein interactions imex curation Sequential tethering of Golgins and catalysis of SNAREpin assembly by the vesicle-tethering protein p115. Sequential tethering of Golgins and catalysis of SNAREpin assembly by the vesicle-tethering protein p115. J. Cell Biol. (0021-9525) 2002 imex curation Shorter J., Beard M.B., Seemann J., Dirac-Svejstrup A.B., Warren G. graham.warren@yale.edu 22-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Shorter J., Beard M.B., Seemann J., Dirac-Svejstrup A.B., Warren G. graham.warren@yale.edu Accepted 2015-JAN-05 by ORCHARD J. Cell Biol. (0021-9525) 2002 Only protein-protein interactions imex curation Sequential tethering of Golgins and catalysis of SNAREpin assembly by the vesicle-tethering protein p115. Sequential tethering of Golgins and catalysis of SNAREpin assembly by the vesicle-tethering protein p115. J. Cell Biol. (0021-9525) 2002 imex curation Shorter J., Beard M.B., Seemann J., Dirac-Svejstrup A.B., Warren G. graham.warren@yale.edu 22-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions rat-liver rattus norvegicus liver Rat liver liver anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Shorter J., Beard M.B., Seemann J., Dirac-Svejstrup A.B., Warren G. Golgi membranes were disassembled with NSF, a vesicle-fusing ATPase, and solubilized with Triton X-100 buffer. SNARE complexes were subsequently allowed to reassemble in the presence of tether proteins. graham.warren@yale.edu Accepted 2015-JAN-05 by ORCHARD J. Cell Biol. (0021-9525) 2002 Only protein-protein interactions imex curation bet1l_rat BET1-like protein Bet1l Gs15 Golgi SNARE with a size of 15 kDa Vesicle transport protein GOS15 protein protein rat Rattus norvegicus (Rat) Rat MADWTRAQSSGAVEEIVDRENKRMADSLASKVTRLKSLALDIDRDTEDQNRYLDGMDSDFTSVTGLLTGSVKRFSTVARSGRDTRKLLCGMAVVLIVAFFILSYLFSRTRT 905EDA73F6AEB4B4 sc22b_rat Vesicle-trafficking protein SEC22b Sec22b SEC22 vesicle-trafficking protein homolog B SEC22 vesicle-trafficking protein-like 1 ER-Golgi SNARE of 24 kDa Sec22l1 protein protein rat Rattus norvegicus (Rat) Rat MVLLTMIARVADGLPLAASMQEDEQSGRDLQQYQSQAKQLFRKLNEQSPTRCTLEAGAMTFHYIIEQGVCYLVLCEAAFPKKLAFAYLEDLHSEFDEQHGKKVPTVSRPYSFIEFDTFIQKTKKLYIDSRARRNLGSINTELQDVQRIMVANIEEVLQRGEALSALDSKANNLSSLSKKYRQDAKYLNMRSTYAKLAAVAVFFIMLIVYVRFWWL 29B4C55961C5A044 scfd1_rat Sec1 family domain-containing protein 1 Syntaxin-binding protein 1-like 2 Vesicle transport-related protein Ra410 SLY1 homolog Scfd1 Ra410 Sly1 Stxbp1l2 protein protein rat Rattus norvegicus (Rat) Rat MVGSKMAASIRERQTVALKRMLNFNVPHVKNSPGEPVWKVLIYDRFGQDIISPLLSVKELRDMGITLHLLLHSDRDPIRDVPAVYFVMPTEENIDRLCQDLRNQLYESYYLNFISAISRSKLEDIANAALAANAVTQVAKVFDQYLNFITLEEDMFVLCNQNKELVSYRAINRPDITDTEMETVMDTIVDSLFCFFVTLGAVPIIRCSRGTAAEMVAVKLDKKLRENLRDARNSLFTGDPLGTGQFSFQRPLLVLVDRNIDLATPLHHTWTYQALVHDVLDFHLNRVNLEESTGVENSPTGARPKRKNKKSYDLTPVDKFWQKHKGSPFPEVAESVQQELESYRAQEDEVKRLKSIMGLEGEDEGAISMLSDNTAKLTSAVSSLPELLEKKRLIDLHTNVATAVLEHIKARKLDVYFEYEEKIMSKTTLDKSLLDVISDPDAGTPEDKMRLFLIYYISAQQAPSEVDLEQYKKALTDAGCNLSPLQYIKQWKAFAKMASTPASYGNTTTKPMGLLSRVMNTGSQFVMEGVKNLVLKQQNLPVTRILDNLMEMKSNPETDDYRYFDPKMLRSNDSSVPRNKSPFQEAIVFVVGGGNYIEYQNLVDYIKGKQGKHILYGCSEIFNATQFIKQLSQLGQK 9ABFBBEBD2862341 gosr1_rat Golgi SNAP receptor complex member 1 Gosr1 Gs28 28 kDa Golgi SNARE protein 28 kDa cis-Golgi SNARE p28 protein protein rat Rattus norvegicus (Rat) Rat MAAGTSNYWEDLRKQARQLENELDLKLVSFSKLCTSYSHSSARDGGRDRYSSDTTPLLNGSSQDRMFETMAIEIEQLLARLTGVNDKMAEYTHSAGVPSLNAALMHTLQRHRDILQDYTHEFHKTKANFMAIRERENLMGSVRKDIESYKSGSGVNNRRTELFLKEHDHLRNSDRLIEETISIAMATKENMTSQRGMLKSIHSKMNTLANRFPAVNSLIQRINLRKRRDSLILGGVIGICTILLLLYAFH mint Involved in transport from the ER to the Golgi apparatus as well as in intra-Golgi transport 0F85EDCE7ADA32E2 stx5_rat Syntaxin-5 Stx5 Stx5a protein protein rat Rattus norvegicus (Rat) Rat MIPRKRYGSKNTDQGVYLGLSKTQVLSPATAINSSSDIAPLPTPVALVPSPPDTMSCRDRTQEFLSACKSLQSRQNGIQTNKPALHATRQCSEFTLMARRIGKDLSNTFAKLEKLTILAKRKSLFDDKAVEIEELTYIIKQDINSLNKQIAQLQDFVRAKGSQSGRHLQTHSNTIVVSLQSKLASMSNDFKSVLEVRTENLKQQRNRREQFSRAPVSALPLAPNNLGGGPIVLGGESRASRDVAIDMMDPRTSQQLQLIDEQDSYIQSRADTMQNIESTIVELGSIFQQLAHMVKEQEETIQRIDENVLGAQLDVEAAHSEILKYFQSVTSNRWLMVKIFLILIVFFIIFVVFLA 0B0D0B2C261A39B4 q63714_rat Golgb1 protein protein rat Rattus norvegicus (Rat) Rat MLSRLSGLANNVLHELSGDYTGWTTMTSSEAELPQGSAMELNNSTQKDVLDRLADAEKLVLELKDIISKQDAQLQQKDEALQEEKKGAESKIKKIKLHAKAKITSLNKQIEELKTQGGAASPPEVQAEELSKHDKSSTEEEMEVEKIKHELQEKEKLISNLQAQLDQAQSEQASQLDKSSAEMEDFILMRQKLQEKEELIGALQTQLSQTQAEQAAQKLRVMQRKLEEHEEALLGRAQVVDLLQKELTSAEQRNQDLSQQLQLLEAEHSTLRNTMEAERQESKILMEKVELEMAERKEELYQLQGQLERAGQAQAELEMQYGTLQQRHETEMEEKTACISLLQKNEQELQSACDALKEENSKLLQEQQEQAAKSAQALQQLEDELQQKSKEISQFVNKPNLEKHETSSQTSLPDVYNEGVQAVMEESVASLQKRVLELENEKGALLLSSLELEELRAENEKLCSRITLLEAQNRAGEADGMVCEVSTAGIALLNRSDSSTEESGQDVLENTFSQKHKELSVLLVEMKEAQEEIAFLKSQLQGKRPEGDYEVLDRKEVQMMESEGLPSVTARDVLCAPRDKNSVPAVEGEQAGMRDQHGTLEAGPLNDTGMELNSPQPDGVDKSLSAPHVCLCHQGELERLKTQVLELETSLHTAKETHEKNLSEKAKEISSLTQLTKEFKESAEEARSTLTAVCEERDQLLYRVKELDVLGELRAQVRELETSLAEAEKQRGLDYESQRAQHNLLTEQIHSLSIEAKSKDVKIETLQRELDGVQLQFSEQGTQIKSLQSQLQTKESEVLEGAERMKDISKEMEELSQALSQKELEIAQMDQLLLEKKKDVETLQQTIQEKDQQVTELSFSMTEKMVQLNEEKFSLGVEIKTLKEQLSLLSRAEGAKREQVEDSGAESSPKHGPHESSAEEPVCKEALQQELEWLRKESEQRKRKLQAALISRKELLQKVSKLEEELAKVREESTKDSLRESEKRELEEDSKNKDDPEKYGTSEWRELEVSLRLTISEKEVELEGIRRDLKEKAAAEEELQALVQRMTQDLQNKTKQIDLLQEEITENQATIQKFITGTMDAGDGDSAVKETSVSSPPRAGGGEHWKPELEGKIVDLEKEKTQLQKKLQEALISRKAILKKAQEKEKHLKEELKEQKDAYRHLQEQFDGQSKENENIRAPLRQLQAKESTDQQLPGTGQQEPTHGSEGLSLEGTEPASESDLHAAQPSHPGETATLQATVSVAQIQDQLKEIEVEKEELELKISSTTSELTKKSEEVLLLQEQINEQGLEIQNLKAASHEAKAHTEQLKQELESSQLKIADLEHLKTLQPELETLQKHVGQKEEEVSYLVGQLGEKEQTLTTVQTEMEEQERLIKALHTQLEMQAKEHEERLKQVQVEICELKKQPKELEEESKAKQQLQRKLQAALISRKEALKENKSLQEQLSSARDAVEHLTKSLADVESQVSVQNQEKDALLGKLALLQEERDKLIVEMDKSLLENQSLGGSCESLKLALGGLTEDKEKLMKELESVRCSKIAESTEWQEKHKELQKEYEVLLQSYENVSNEAERIQHVVESVRQEKQEVYAKLRSAESDKREREKQLQDAEQEMEEMKEKMRKFAKSKQQKILELEEENDRLRAEAQPVGGANESMEALLSSNASLKEELERITLEYKTLSKEFEALMAEKNTLSEETRNLKLQVEAQELKQASLETTEKSDEPKDVIEEVTEAVVGKSQEQDSLSENAKLEDAEATLLANSAKPGVSETFSSHDDINNYLQQLDQLKGRIAELEMEKQKDRELSQTLENEKNALLTQISAKDSELKLLEEEVAKINMLNQQIQEELSRVTKLKETAEEEKDDLEERLMNQLAELNGSIGNYYQDVTDAQIKNEQLESEMQNLKRCVSELEEEKQQLVKEKTKVESEIRKEYMEKIQGAQKGPGSKIHAKELQELLKEKQQEVKQLQKDCIRYLGRISALEKTVKALEFVHTESQKDLDATKGNLAQAVEHHKKAQAELSSFKILLDDTQSEAARVLADNLKLKKELQSNKESIKSQIKQKDEDLLRRLEQAEEKHRKEKKNMQEKLDALHREKAHVEDTLAEIQVSLTRKDKDMKELQQSLDSTLAQLAAFTKSMSSLQDDRDRVIDEAKKWEQRFGDAIQTKEEEVRLKEENCTALKDQLRQMTIHMEELKITVSRLEHDKEIWESKAQTELQHQQKAYDKLQEENKELMSQLEEAGQLYHDSKNELTKLESELKSLKDQSTDLKNSLEKCREHENNLEGIIKQQEADIQNCKFNCEQLETDLTASRELTTRLHDEINVKEQKIISLLSGKEEAIQVAIAELHQQHSKEIKELENLLSQEEEENLTLEEENKRAVEKTNQLTEALETIKKESLEQKAQLDSFVKSMSSLQDDRDRIVSDYRQLEERHLSVILEKDELIQDAAAENNKLKEEIRGLRGHMDDLNSENAKLDAELIQYRRDLNEVITIKDSQQRQLLEAQLQQNKELRNECVKLEGRLKGSEAEKQSLQMSLDALQEENQGLSKEIKSFKEQLTALHEEGALAVYHAQLRVREEEVQKLTAALSSSQKRTVDLQEELVCVQKEASKKVSEIEDKLKRELKHLHHNAGIMRNETETAEERVAELARDLVEMEQKLLTVTKENKDLTAQIQAFGKSMSSLQDSRDHATEELSDLKKKYDASLKELAQLKGRQDLGRESDVLSQAAFPLTTSENISSRLEKLNQQLISKDEQLLHLSSELESSHNQVQSFTKAMTSLQNERDHLWNELEKFRKSEEGKQRSAAPSAASSPAEVQSLKKAMSSLQNDRDRLLKELKNLQQQYLQMNQEITELRPLKAQLQESQDQTKALQMMKEELRQENLSWQHELDQLRVEKNSWELHERRMKEQYLMAISDKDQQLGHLQNLLRELRSSSQTQILPTQYQRQASSGTSAALDGSQNLVYETDCLRTQLNDSLKEIHQKELRIQQLNSKFSQLLEEKNILSTQLRDANQSLRDSQHHYSNLFNHCAILERQVQQLQATGPLNADVAPGAPQEKDGIHIKTETETTGEEQPSFSEVQQQLCNTKHDLSELKKLLEEERDQRLTAENALSLAKEQIRRLEHSEWESARTPIIGACGSQEQVLLMDLPGSSCRRTRSSAGWKRVLRSLCRSRTRVPLLAAIYFLMIHVLLVLCFTGHL mint 48F035DF43647F51 bet1_rat BET1 homolog Bet1 Golgi vesicular membrane-trafficking protein p18 protein protein rat Rattus norvegicus (Rat) Rat MRRAGLGDGAPPGGYGNYGYANSGYNACEEENDRLTESLRSKVTAIKSLSIEIGHEVKNQNKLLAEMDSQFDSTTGFLGKTMGRLKILSRGSQTKLLCYMMLFSLFVFFVIYWIIKLR 9ED4863140DAF051 uso1_rat General vesicular transport factor p115 Uso1 Vdp Protein USO1 homolog Transcytosis-associated protein Vesicle-docking protein protein protein rat Rattus norvegicus (Rat) Rat MNFLRGVMGGQSAGPQHTEAETIQKLCDRVASSTLLDDRRNAVRALKSLSKKYRLEVGIQAMEHLIHVLQTDRSDSEIIAYALDTLYNIISNDEEEEVEENSTRQSEDLGSQFTEIFIKQPENVTLLLSLLEEFDFHVRWPGVRLLTSLLKQLGPPVQQIILVSPMGVSKLMDLLADSREIIRNDGVLLLQALTRSNGAIQKIVAFENAFERLLDIITEEGNSDGGIVVEDCLILLQNLLKNNNSNQNFFKEGSYIQRMKAWFEVGDENPGWSAQKVTNLHLMLQLVRVLVSPTNPPGATSSCQKAMFQCGLLQQLCTILMATGIPADILTETINTVSEVIRGCQVNQDYFASVNAPSNPPRPAIVVLLMSMVNERQPFVLRCAVLYCFQCFLYKNEKGQGEIVATLLPSTIDATGNSVSAGQLLCGGLFSTDSLSNWCAAVALAHALQGNATQKEQLLRVQLATSIGNPPVSLLQQCTNILSQGSKIQTRVGLLMLLCTWLSNCPIAVTHFLHNSANVPFLTGQIAENLGEEEQLVQGLCALLLGISIYFNDNSLENYTKEKLKQLIEKRIGKENYIEKLGFISKHELYSRASQKPQPNFPSPEYMIFDHEFTKLVKELEGVITKAIYKSSEEDKKEEEVKKTLEQHDNIVTHYKNMIREQDLQLEELKQQVSTLKCQNEQLQTAVTQQASQIQQHKDQYNLLKVQLGKDNHHQGSHSDGAQVNGIQPEEISRLREEIEELRSHQVLLQSQLAEKDTVIENLRSSQVSGMSEQALATCSPRDAEQVAELKQELSALKSQLCSQSLEITRLQTENSELQQRAETLAKSVPVEGESELVTAAKTTDVEGRLSALLQETKELKNEIKALSEERTAIQKQLDSSNSTIAILQTEKDKLYLEVTDSKKEQDDLLVLLADQDQKILSLKSKLKDLGHPVEEEDESGDQEDDDDELDDGDRDQDI 356394B48C7E003B gosr2_rat Golgi SNAP receptor complex member 2 Gosr2 Gs27 27 kDa Golgi SNARE protein Membrin protein protein rat Rattus norvegicus (Rat) Rat MEPLYQQTHKQVHEIQSHMGRLETADKQSVHLVENEIQASIDQIFSHLERLEILSSKEPPNRRQNAKLRVDQLKYDVQHLQTALRNFQHRRQAKEQQERQRDELLSRTFTTNDSDTTIPMDESLQFNSSLQNIHHGMDDLIGGGHSILEGLRAQRLTLKGTQKKILDIANMLGLSNTVMRLIEKRAFQDKYFMIGGMLLTCAVMFLVVQYLT 164380B16A4532A0 goga2_rat Golgin subfamily A member 2 Golga2 130 kDa cis-Golgi matrix protein protein protein rat Rattus norvegicus (Rat) Rat MWPPRFPPPRPGMSEETRQSKLAAAKKKLREYQQKNSPGVPAGAKKKKKIKNGHSPERTSASDCQSAENVPTDHTAPAPPSTAAATMFLGVVPSPDADLIQSHDAGNCSNLMEETKTFSSTESLRQLSQQLNGLVSESTSYINGEGLTSSNMKELESRYQELAVALDSSYVTNKQLSSTIEELKQQNQDTLDQLEKEKKDYQQKLAKEQGALREQLQVHIQTIGILVSEKAELQTALAHTQQAARQKAGESEDLASRLQSSRQRVGELERTLSTVSTQQKQADRYNKDLTKERDALKLELYKNGKSNEDLRQQNSELEEKLRVLVAEKAAAQLGAEELQKKLEMSELLLQQFSSQSEASGSNEQLQQAMEERAQLESHVGQLMESLKQLQVERDQYAENLKGESAMWQQRVQQMAEQVHALKEEKEQRESQVQELEASLAELRSQMEEPPPPEPPTGPSEAEERLQGEVEQLQKELEGLTGQLRAQVQDNESLSHLNREQEGRLLELEREAQHWSEQAEERKQILESMQSDRTTISRALSQNRELKEQLAELQNGFVRLTNENMEITSALQSEQHVKKELARKLGELQERLGELKETVELKSQEAQGLQEQRDQCLSHLQQYAAAYQQHLTAYEQLTSEKEALHKQLLLQTQLMDQLQHEEVQGKMAAEMARQELQEAQERLKASSQENQQLQAQLSLLVLPGEDMDQEEQDEEVPQPSLTIPEDLVSREAMVAFCNAAIARAEEEQARLRVQLREQKARCRSLAHLAAPVQSKLEKEAVVPRDMGDSVSEESNQALHVAMEKLQNRFLEVMQEKVELKERVEELEHCCIQLSGETDTIGEYIALYQNQRAVLKARHLEKEEYISRLAQDKEEMKVKLLELQELVLRLVNERNEWQGKFLAVSQNPADVPAPVPTGSQEFGAADQQGDLREVSLADDTEPAQGEAGVPAPQENPTAQQIMQLLREIQNPQERPGLGSNPCIPFFYRADENDEVKIMVI 3AA8CBC61F98A6A4 vti1a_rat Vesicle transport through interaction with t-SNAREs homolog 1A Vti1a Vti1l2 Vesicle transport v-SNARE protein Vti1-like 2 Vti1-rp2 protein protein rat Rattus norvegicus (Rat) Rat MSADFEGYEQDFAVLTAEITSKISRVPRLPPDEKKQMVANVEKQLEEARELLEQMDLEVREIPPQSRGMYSNRMRSYKQEMGKLETDFKRSRIAYSDEVRNELLGDAGNSSENQLIKLREERAHLLDNTERLERSSRRLEAGYQIAVETEQIGQEMLENLSHDRERIQRARERLRETDANLGKSSRILTGMLRRIIQNRILLVILGIIVVITILTAITFFVRGH mint V-SNARE that mediates vesicle transport pathways through interactions with t-SNAREs on the target membrane. These interactions are proposed to mediate aspects of the specificity of vesicle trafficking and to promote fusion of the lipid bilayers. May be required for a fusion step during synaptic vesicle Recycling in the nerve terminal 508CAC274ED055C4 ykt6_rat Synaptobrevin homolog YKT6 Ykt6 protein protein rat Rattus norvegicus (Rat) Rat MKLYSLSVFYKGEPKAVLLKAAYDVSSFSFFQRSSVQEFMTFTSQLIVERSAKGSRASVKEQEYLCHVYVRSDSLAGVVIADSEYPSRVAFTLLEKVLDEFSKQVDRIDWPVGSPATIHYTALDGHLSRYQNPREADPMSKVQAELDETKIILHNTMESLLERGEKLDDLVSKSEVLGTQSKAFYKTARKQNSCCAIM 384F4EDBC8295407 uso1-bet1 3 10 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 8 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 12 unspecified role unspecified role bait bait biotin tag biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined rat-liver rattus norvegicus liver Rat liver liver 5 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 11 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 6 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 14 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 13 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 16 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 9 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 7 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association Fig. 1c Instead of protein rSec22p the mammalian Sec22b was used as the former is the yeast homologue. In addition to the Western Blot a silver staining was also performed on another SDS-PAGE gel (Fig. 1b) but only p115, giantin and GM130 were identified by weight (in p115 pulldown). uso1-bet1-1 3 8 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 6 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 13 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 9 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 11 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 16 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 5 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 7 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 12 unspecified role unspecified role bait bait cc1 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain biotin tag biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Fig. 1c Instead of protein rSec22p the mammalian Sec22b was used as the former is the yeast homologue. Bet1 and GS15 (Bet1l) bands were significantly weaker than with the FL p115 (Uso1) pulldown. In addition to the Western Blot a silver staining was also performed on another SDS-PAGE gel (Fig. 1b) but only p115, giantin and GM130 were identified by weight (in p115 pulldown). uso1-bet1-2 3 8 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 11 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 13 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 6 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 16 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 5 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 12 unspecified role unspecified role bait bait cc4 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain biotin tag biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Fig. 1c Instead of protein rSec22p the mammalian Sec22b was used as the former is the yeast homologue. Bet1 and GS15 (Bet1l) bands were significantly weaker than with the FL p115 (Uso1) pulldown. In addition to the Western Blot a silver staining was also performed on another SDS-PAGE gel (Fig. 1b) but only p115, giantin and GM130 were identified by weight (in p115 pulldown). uso1-bet1-3 3 10 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 8 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 14 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 13 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 6 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 9 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 5 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 12 unspecified role unspecified role bait bait tail-acidic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain his tag 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 rat-liver rattus norvegicus liver Rat liver liver 11 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 16 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 7 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association Fig. 1c Instead of protein rSec22p the mammalian Sec22b was used as the former is the yeast homologue. Bet1 and GS15 (Bet1l) bands were weaker than for the FL p115 (Uso1) pulldown. In addition to the Western Blot a silver staining was also performed on another SDS-PAGE gel (Fig. 1b) but only p115, giantin and GM130 were identified by weight (in p115 pulldown). uso1-bet1-4 3 8 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 11 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 16 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 5 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 12 unspecified role unspecified role bait bait his tag 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 tail domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain rat-liver rattus norvegicus liver Rat liver liver 7 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 14 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 9 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 6 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 13 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association Fig. 1c Instead of protein rSec22p the mammalian Sec22b was used as the former is the yeast homologue. Bet1 and GS15 (Bet1l) bands were weaker than for the FL p115 (Uso1) pulldown. In addition to the Western Blot a silver staining was also performed on another SDS-PAGE gel (Fig. 1b) but only p115, giantin and GM130 were identified by weight (in p115 pulldown). gosr1-bet1-1 4 8 unspecified role unspecified role bait bait 14 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 16 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey physical association physical association Fig. 2a (lane 2), 2c gosr1-bet1-4 4 8 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 14 unspecified role unspecified role prey prey 16 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 12 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association Fig. 2a (lanes 3-6), 2c When p115 is included, the SNARE bands are stronger and Giantin only starts to bind at 20 nM p115. stx5-bet1-6 4 15 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 13 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 16 unspecified role unspecified role prey prey 14 unspecified role unspecified role prey prey 12 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association When p115 is included, the SNARE bands are stronger. Vti1a bands are very faint. Fig. 2b (lanes 3-6), 2d stx5-bet1-2 4 8 unspecified role unspecified role prey prey 15 unspecified role unspecified role prey prey 14 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey 16 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 13 unspecified role unspecified role prey prey physical association physical association Fig. 2b (lane 2), 2d Vti1a bands are very faint. gosr1-bet1-3 4 8 unspecified role unspecified role bait bait 16 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey physical association physical association Fig. 2e (lane 2) stx5-bet1-5 4 8 unspecified role unspecified role prey prey 13 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 16 unspecified role unspecified role prey prey physical association physical association Fig. 2f (lane 2) bet1l-gosr1-3 4 8 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait 16 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 12 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association Fig. 3a (lanes 4-6, 10-11, 13) Addition of p115 CC2 domain peptide (lanes 10 & 13) had no effect on the interaction. gosr2-bet1 4 13 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey physical association physical association Fig. 3c (lanes 3, 7-9, 12, 14), 3d Combined addition of p115 and its CC1 domain peptide (3c, lane 8) and a wash with buffer containing the CC1 domain peptide (3a, lane 12) or 1M KCL (3a, lane 14) abolished the binding of p115 to the complex. Boiling of the re-constituted complexes destroyed them completely (3d). Addition of p115 CC2 domain peptide (3c, lane 9) had no effect on the interaction. p115 (Uso1, P41542), either in reconstitution butter or in wash buffer. CC1 domain (637-699) of p115 (Uso1, P41542) 1M KCl wash stx5-uso1 1 12 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 9 unspecified role unspecified role bait bait cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range his tag 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 6a (top) Addition of CC1 peptide reduced the p115 band but not the syntaxin-5 band. Addition of CC2 or CC3 domain peptides made no difference to the interaction. CC1 domain (637-699) of p115 (Uso1, P41542) gosr1-uso1-1 1 8 unspecified role unspecified role bait bait cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range his tag 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 ecoli Escherichia coli (strain K12) 12 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association Fig. 6a (bottom), 6d Addition of CC1 peptide reduced the p115 band but not the syntaxin-5 band. Addition of CC2 or CC3 domain peptides made no difference to the interaction. CC1 domain (637-699) of p115 (Uso1, P41542) gosr1-uso1-2 2 8 unspecified role unspecified role bait bait his tag 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 cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range ecoli Escherichia coli (strain K12) 12 unspecified role unspecified role prey prey tail domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain rat-liver rattus norvegicus liver Rat liver liver 157 155 physical association physical association Fig. 6b (top left) stx5-uso1-1 1 9 unspecified role unspecified role bait bait cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range gst tag 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) 12 unspecified role unspecified role prey prey tail domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain rat-liver rattus norvegicus liver Rat liver liver 160 163 physical association physical association Fig. 6b (bottom left), 6c gosr1-bet1-5 4 8 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey 12 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 9 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 16 unspecified role unspecified role prey prey physical association physical association Fig. 2e (lanes 3, 5-7) The addition of CC2 domain (728-765) of p115 (Uso1, P41542), C-terminal domain (521-595) of p115 (Uso1, P41542) or N-terminal domain (1-73) of GM130 (Golga2, Q62839) had not effect on the interaction. gosr1-bet1-6 4 8 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey 16 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey physical association physical association Fig. 2e (lane 4) CC1 domain (637-699) of p115 (Uso1, P41542) p115 (Uso1, P41542) The combination of adding full length p115 and its CC1 domain peptide results in weaker bands of all SNAREs and a lack of detection of p115 by WB while p115 on its own enhances the SNARE interactions (see EBI-10020539). stx5-bet1-7 4 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 16 unspecified role unspecified role prey prey 13 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey physical association physical association Fig. 2f (lane 4) p115 (Uso1, P41542) CC1 domain (637-699) of p115 (Uso1, P41542) The combination of adding full length p115 and its CC1 domain peptide results in weaker bands of all SNAREs and a lack of detection of p115 by WB while p115 on its own enhances the SNARE interactions (see EBI-10020539). stx5-bet1-8 4 8 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 13 unspecified role unspecified role prey prey 16 unspecified role unspecified role prey prey physical association physical association Fig. 2f (lanes 3, 5-7) The addition of CC2 domain (728-765) of p115 (Uso1, P41542), C-terminal domain (521-595) of p115 (Uso1, P41542) or N-terminal domain (1-73) of GM130 (Golga2, Q62839) had not effect on the interaction. bet1l-gosr1-2 4 8 unspecified role unspecified role prey prey 16 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association Fig. 3a (lanes 3, 7-9, 12, 14), 3b, 8e, 8f CC1 domain (637-699) of p115 (Uso1, P41542), either in reconstitution butter or in wash buffer. p115 (Uso1, P41542) Combined addition of p115 and its CC1 domain peptide (3a, lane 8) and a wash with buffer containing the CC1 domain peptide (3a, lane 12) or 1M KCL (3a, lane 14) abolished the binding of p115 to the complex. Boiling of the re-constituted complexes destroys them completely (3b). Addition of p115 CC2 domain peptide (3a lane 9 & 3b lane 7) had no effect on the interaction. 1M KCl wash gosr2-bet1-1 4 13 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 12 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association Fig. 3c (lanes 4-6, 10-11, 13) Addition of p115 CC2 domain peptide (lanes 10 & 13) had no effect on the interaction. giantin-golga2 4 10 unspecified role unspecified role bait bait 14 unspecified role unspecified role prey prey 12 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association Fig. 5a GM130 NT domain (GOLGA2, Q62839) p115 (Uso1, P41542) Addition of GM130 NT domain (GOLGA2, Q62839) produced only very faint bands for GM130 full length and p115. stx5-gosr1 1 8 unspecified role unspecified role prey prey his tag 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 cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range ecoli Escherichia coli (strain K12) 12 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver 9 unspecified role unspecified role bait bait gst tag 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 cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range ecoli Escherichia coli (strain K12) physical association physical association Fig. 7a (top panel, lanes 5-10), 7b (top panel lanes 9, 11-13), 7d, 8a (top panel lanes 5 & 6, middle panel lanes 5 & 6, bottom panel lanes 5 & 6), 8b (left & right panels) Increasing amounts of p115 increase the binding of GOS28 (7a, 8a). Addition of p115 CC2 or CT domains, or GM130 NT domain had no effect on the interaction (7b, lanes 11-13, 8a bottom panel, 8b right panel). gosr1-stx5 1 8 unspecified role unspecified role bait bait cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range his tag 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 ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role prey prey cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range gst tag 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) 12 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association Fig. 7a (bottom panel, lanes 5-10), 7b (bottom, panel lanes 9-13) p115 (Uso1) is not required for efficient binding of GOS28 and syntaxin-5 (7a). Addition of p115 CC1, CC2 or CT domains, or GM130 NT domain had no effect on the interaction (7b, lanes 11-13). Addition of p115 CC1 domain reduced the binding of p115 to the complex. gosr1-stx5-1 1 8 unspecified role unspecified role bait bait his tag 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 cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role prey prey gst tag 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 cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range ecoli Escherichia coli (strain K12) 227 230 physical association physical association Fig. 7a (bottom panel, lane 4), 7b (bottom panel, lanes 4-8). Addition of p115 CC1, CC2 or CT domains, or GM130 NT domain had no effect on the interaction (7b, lanes 5-8). stx5-gosr1-1 1 8 unspecified role unspecified role prey prey his tag 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 cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role bait bait cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range gst tag 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) 236 234 physical association physical association Fig. 7a (top panel, lane 4), 7b (top panel lanes 4-8), 7c, 8a (top panel lanes 4, 9-11, middle panel lanes 4, 9-11), 8b (middle panel), 8c (lanes 4 & 5), 8d Binding of GOS28 and syntaxin-5 is weak without the addition of p115 (7a lane 4, 7b lanes 4-8, 8a all panels, 8d). Addition of p115 CC1, CC2 or CT domains, or GM130 NT domain had no effect on the interaction (7b, lanes 5-8, 8a bottom panel). Addition of p115 CC1 domain peptides (either in binding or wash buffer) with full length p115 disrupted the binding of p115 to the complex (7b, lane 10, 8a middle panel lanes 10-11, 8b middle panel). Aggitation also disrupted the binding of p115 to the complex (8c lane 5). gosr1-stx5-2 1 8 unspecified role unspecified role unspecified role unspecified role cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range his tag 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 ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role unspecified role unspecified role cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range gst tag 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) 240 243 physical association physical association Fig. 7e Addition of increasing amounts of p115 head, tail (T, 637-930 aa) and tail plus acidic region (TA, 637-959 aa) increase the interaction of GOS28-syntaxin-5 but >1500 nM of T or TA constructs reduce the binding efficacy. Bait/prey relationship is unknown.