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/ http://www.ebi.ac.uk/ shorter-2002-2 Sequential tethering of Golgins and catalysis of SNAREpin assembly by the vesicle-tethering protein p115. 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 J. Cell Biol. (0021-9525) 2002 Only protein-protein interactions imex curation shorter-2002-3 Sequential tethering of Golgins and catalysis of SNAREpin assembly by the vesicle-tethering protein p115. 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 J. Cell Biol. (0021-9525) 2002 Only protein-protein interactions imex curation shorter-2002-1 Sequential tethering of Golgins and catalysis of SNAREpin assembly by the vesicle-tethering protein p115. 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 J. Cell Biol. (0021-9525) 2002 Only protein-protein interactions imex curation shorter-2002-4 Sequential tethering of Golgins and catalysis of SNAREpin assembly by the vesicle-tethering protein p115. 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 J. Cell Biol. (0021-9525) 2002 Only protein-protein interactions imex curation 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 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 ykt6_rat Synaptobrevin homolog YKT6 Ykt6 protein protein rat Rattus norvegicus (Rat) Rat MKLYSLSVFYKGEPKAVLLKAAYDVSSFSFFQRSSVQEFMTFTSQLIVERSAKGSRASVKEQEYLCHVYVRSDSLAGVVIADSEYPSRVAFTLLEKVLDEFSKQVDRIDWPVGSPATIHYTALDGHLSRYQNPREADPMSKVQAELDETKIILHNTMESLLERGEKLDDLVSKSEVLGTQSKAFYKTARKQNSCCAIM stx5_rat Syntaxin-5 Stx5 Stx5a protein protein rat Rattus norvegicus (Rat) Rat MIPRKRYGSKNTDQGVYLGLSKTQVLSPATAINSSSDIAPLPTPVALVPSPPDTMSCRDRTQEFLSACKSLQSRQNGIQTNKPALHATRQCSEFTLMARRIGKDLSNTFAKLEKLTILAKRKSLFDDKAVEIEELTYIIKQDINSLNKQIAQLQDFVRAKGSQSGRHLQTHSNTIVVSLQSKLASMSNDFKSVLEVRTENLKQQRNRREQFSRAPVSALPLAPNNLGGGPIVLGGESRASRDVAIDMMDPRTSQQLQLIDEQDSYIQSRADTMQNIESTIVELGSIFQQLAHMVKEQEETIQRIDENVLGAQLDVEAAHSEILKYFQSVTSNRWLMVKIFLILIVFFIIFVVFLA 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 gosr2_rat Golgi SNAP receptor complex member 2 Gosr2 Gs27 27 kDa Golgi SNARE protein Membrin protein protein rat Rattus norvegicus (Rat) Rat MEPLYQQTHKQVHEIQSHMGRLETADKQSVHLVENEIQASIDQIFSHLERLEILSSKEPPNRRQNAKLRVDQLKYDVQHLQTALRNFQHRRQAKEQQERQRDELLSRTFTTNDSDTTIPMDESLQFNSSLQNIHHGMDDLIGGGHSILEGLRAQRLTLKGTQKKILDIANMLGLSNTVMRLIEKRAFQDKYFMIGGMLLTCAVMFLVVQYLT 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 bet1_rat BET1 homolog Bet1 Golgi vesicular membrane-trafficking protein p18 protein protein rat Rattus norvegicus (Rat) Rat MRRAGLGDGAPPGGYGNYGYANSGYNACEEENDRLTESLRSKVTAIKSLSIEIGHEVKNQNKLLAEMDSQFDSTTGFLGKTMGRLKILSRGSQTKLLCYMMLFSLFVFFVIYWIIKLR q63714_rat Golgb1 protein protein rat Rattus norvegicus (Rat) Rat MLSRLSGLANNVLHELSGDYTGWTTMTSSEAELPQGSAMELNNSTQKDVLDRLADAEKLVLELKDIISKQDAQLQQKDEALQEEKKGAESKIKKIKLHAKAKITSLNKQIEELKTQGGAASPPEVQAEELSKHDKSSTEEEMEVEKIKHELQEKEKLISNLQAQLDQAQSEQASQLDKSSAEMEDFILMRQKLQEKEELIGALQTQLSQTQAEQAAQKLRVMQRKLEEHEEALLGRAQVVDLLQKELTSAEQRNQDLSQQLQLLEAEHSTLRNTMEAERQESKILMEKVELEMAERKEELYQLQGQLERAGQAQAELEMQYGTLQQRHETEMEEKTACISLLQKNEQELQSACDALKEENSKLLQEQQEQAAKSAQALQQLEDELQQKSKEISQFVNKPNLEKHETSSQTSLPDVYNEGVQAVMEESVASLQKRVLELENEKGALLLSSLELEELRAENEKLCSRITLLEAQNRAGEADGMVCEVSTAGIALLNRSDSSTEESGQDVLENTFSQKHKELSVLLVEMKEAQEEIAFLKSQLQGKRPEGDYEVLDRKEVQMMESEGLPSVTARDVLCAPRDKNSVPAVEGEQAGMRDQHGTLEAGPLNDTGMELNSPQPDGVDKSLSAPHVCLCHQGELERLKTQVLELETSLHTAKETHEKNLSEKAKEISSLTQLTKEFKESAEEARSTLTAVCEERDQLLYRVKELDVLGELRAQVRELETSLAEAEKQRGLDYESQRAQHNLLTEQIHSLSIEAKSKDVKIETLQRELDGVQLQFSEQGTQIKSLQSQLQTKESEVLEGAERMKDISKEMEELSQALSQKELEIAQMDQLLLEKKKDVETLQQTIQEKDQQVTELSFSMTEKMVQLNEEKFSLGVEIKTLKEQLSLLSRAEGAKREQVEDSGAESSPKHGPHESSAEEPVCKEALQQELEWLRKESEQRKRKLQAALISRKELLQKVSKLEEELAKVREESTKDSLRESEKRELEEDSKNKDDPEKYGTSEWRELEVSLRLTISEKEVELEGIRRDLKEKAAAEEELQALVQRMTQDLQNKTKQIDLLQEEITENQATIQKFITGTMDAGDGDSAVKETSVSSPPRAGGGEHWKPELEGKIVDLEKEKTQLQKKLQEALISRKAILKKAQEKEKHLKEELKEQKDAYRHLQEQFDGQSKENENIRAPLRQLQAKESTDQQLPGTGQQEPTHGSEGLSLEGTEPASESDLHAAQPSHPGETATLQATVSVAQIQDQLKEIEVEKEELELKISSTTSELTKKSEEVLLLQEQINEQGLEIQNLKAASHEAKAHTEQLKQELESSQLKIADLEHLKTLQPELETLQKHVGQKEEEVSYLVGQLGEKEQTLTTVQTEMEEQERLIKALHTQLEMQAKEHEERLKQVQVEICELKKQPKELEEESKAKQQLQRKLQAALISRKEALKENKSLQEQLSSARDAVEHLTKSLADVESQVSVQNQEKDALLGKLALLQEERDKLIVEMDKSLLENQSLGGSCESLKLALGGLTEDKEKLMKELESVRCSKIAESTEWQEKHKELQKEYEVLLQSYENVSNEAERIQHVVESVRQEKQEVYAKLRSAESDKREREKQLQDAEQEMEEMKEKMRKFAKSKQQKILELEEENDRLRAEAQPVGGANESMEALLSSNASLKEELERITLEYKTLSKEFEALMAEKNTLSEETRNLKLQVEAQELKQASLETTEKSDEPKDVIEEVTEAVVGKSQEQDSLSENAKLEDAEATLLANSAKPGVSETFSSHDDINNYLQQLDQLKGRIAELEMEKQKDRELSQTLENEKNALLTQISAKDSELKLLEEEVAKINMLNQQIQEELSRVTKLKETAEEEKDDLEERLMNQLAELNGSIGNYYQDVTDAQIKNEQLESEMQNLKRCVSELEEEKQQLVKEKTKVESEIRKEYMEKIQGAQKGPGSKIHAKELQELLKEKQQEVKQLQKDCIRYLGRISALEKTVKALEFVHTESQKDLDATKGNLAQAVEHHKKAQAELSSFKILLDDTQSEAARVLADNLKLKKELQSNKESIKSQIKQKDEDLLRRLEQAEEKHRKEKKNMQEKLDALHREKAHVEDTLAEIQVSLTRKDKDMKELQQSLDSTLAQLAAFTKSMSSLQDDRDRVIDEAKKWEQRFGDAIQTKEEEVRLKEENCTALKDQLRQMTIHMEELKITVSRLEHDKEIWESKAQTELQHQQKAYDKLQEENKELMSQLEEAGQLYHDSKNELTKLESELKSLKDQSTDLKNSLEKCREHENNLEGIIKQQEADIQNCKFNCEQLETDLTASRELTTRLHDEINVKEQKIISLLSGKEEAIQVAIAELHQQHSKEIKELENLLSQEEEENLTLEEENKRAVEKTNQLTEALETIKKESLEQKAQLDSFVKSMSSLQDDRDRIVSDYRQLEERHLSVILEKDELIQDAAAENNKLKEEIRGLRGHMDDLNSENAKLDAELIQYRRDLNEVITIKDSQQRQLLEAQLQQNKELRNECVKLEGRLKGSEAEKQSLQMSLDALQEENQGLSKEIKSFKEQLTALHEEGALAVYHAQLRVREEEVQKLTAALSSSQKRTVDLQEELVCVQKEASKKVSEIEDKLKRELKHLHHNAGIMRNETETAEERVAELARDLVEMEQKLLTVTKENKDLTAQIQAFGKSMSSLQDSRDHATEELSDLKKKYDASLKELAQLKGRQDLGRESDVLSQAAFPLTTSENISSRLEKLNQQLISKDEQLLHLSSELESSHNQVQSFTKAMTSLQNERDHLWNELEKFRKSEEGKQRSAAPSAASSPAEVQSLKKAMSSLQNDRDRLLKELKNLQQQYLQMNQEITELRPLKAQLQESQDQTKALQMMKEELRQENLSWQHELDQLRVEKNSWELHERRMKEQYLMAISDKDQQLGHLQNLLRELRSSSQTQILPTQYQRQASSGTSAALDGSQNLVYETDCLRTQLNDSLKEIHQKELRIQQLNSKFSQLLEEKNILSTQLRDANQSLRDSQHHYSNLFNHCAILERQVQQLQATGPLNADVAPGAPQEKDGIHIKTETETTGEEQPSFSEVQQQLCNTKHDLSELKKLLEEERDQRLTAENALSLAKEQIRRLEHSEWESARTPIIGACGSQEQVLLMDLPGSSCRRTRSSAGWKRVLRSLCRSRTRVPLLAAIYFLMIHVLLVLCFTGHL mint goga2_rat Golgin subfamily A member 2 Golga2 130 kDa cis-Golgi matrix protein protein protein rat Rattus norvegicus (Rat) Rat MWPPRFPPPRPGMSEETRQSKLAAAKKKLREYQQKNSPGVPAGAKKKKKIKNGHSPERTSASDCQSAENVPTDHTAPAPPSTAAATMFLGVVPSPDADLIQSHDAGNCSNLMEETKTFSSTESLRQLSQQLNGLVSESTSYINGEGLTSSNMKELESRYQELAVALDSSYVTNKQLSSTIEELKQQNQDTLDQLEKEKKDYQQKLAKEQGALREQLQVHIQTIGILVSEKAELQTALAHTQQAARQKAGESEDLASRLQSSRQRVGELERTLSTVSTQQKQADRYNKDLTKERDALKLELYKNGKSNEDLRQQNSELEEKLRVLVAEKAAAQLGAEELQKKLEMSELLLQQFSSQSEASGSNEQLQQAMEERAQLESHVGQLMESLKQLQVERDQYAENLKGESAMWQQRVQQMAEQVHALKEEKEQRESQVQELEASLAELRSQMEEPPPPEPPTGPSEAEERLQGEVEQLQKELEGLTGQLRAQVQDNESLSHLNREQEGRLLELEREAQHWSEQAEERKQILESMQSDRTTISRALSQNRELKEQLAELQNGFVRLTNENMEITSALQSEQHVKKELARKLGELQERLGELKETVELKSQEAQGLQEQRDQCLSHLQQYAAAYQQHLTAYEQLTSEKEALHKQLLLQTQLMDQLQHEEVQGKMAAEMARQELQEAQERLKASSQENQQLQAQLSLLVLPGEDMDQEEQDEEVPQPSLTIPEDLVSREAMVAFCNAAIARAEEEQARLRVQLREQKARCRSLAHLAAPVQSKLEKEAVVPRDMGDSVSEESNQALHVAMEKLQNRFLEVMQEKVELKERVEELEHCCIQLSGETDTIGEYIALYQNQRAVLKARHLEKEEYISRLAQDKEEMKVKLLELQELVLRLVNERNEWQGKFLAVSQNPADVPAPVPTGSQEFGAADQQGDLREVSLADDTEPAQGEAGVPAPQENPTAQQIMQLLREIQNPQERPGLGSNPCIPFFYRADENDEVKIMVI 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 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 bet1l-gosr1-3 1764932 n/a 1764936 unspecified role unspecified role prey prey n/a 1764937 unspecified role unspecified role bait bait n/a 1764938 unspecified role unspecified role prey prey n/a 1764939 unspecified role unspecified role prey prey n/a 1764940 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association false false false 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 1764932 n/a 1764941 unspecified role unspecified role bait bait n/a 1764942 unspecified role unspecified role prey prey n/a 1764943 unspecified role unspecified role prey prey n/a 1764939 unspecified role unspecified role prey prey physical association physical association false false false 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 gosr1-bet1-5 1764932 n/a 1764936 unspecified role unspecified role bait bait n/a 1764943 unspecified role unspecified role prey prey n/a 1764940 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764939 unspecified role unspecified role prey prey n/a 1764937 unspecified role unspecified role prey prey n/a 1764938 unspecified role unspecified role prey prey physical association physical association false false false 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 1764932 n/a 1764936 unspecified role unspecified role bait bait n/a 1764937 unspecified role unspecified role prey prey n/a 1764938 unspecified role unspecified role prey prey n/a 1764943 unspecified role unspecified role prey prey n/a 1764939 unspecified role unspecified role prey prey physical association physical association false false false 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 1764932 n/a 1764936 unspecified role unspecified role prey prey n/a 1764939 unspecified role unspecified role bait bait n/a 1764938 unspecified role unspecified role prey prey n/a 1764941 unspecified role unspecified role prey prey n/a 1764942 unspecified role unspecified role prey prey n/a 1764943 unspecified role unspecified role prey prey n/a 1764937 unspecified role unspecified role prey prey physical association physical association false false false 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 1764932 n/a 1764936 unspecified role unspecified role prey prey n/a 1764937 unspecified role unspecified role prey prey n/a 1764943 unspecified role unspecified role prey prey n/a 1764942 unspecified role unspecified role prey prey n/a 1764939 unspecified role unspecified role bait bait n/a 1764941 unspecified role unspecified role prey prey n/a 1764938 unspecified role unspecified role prey prey physical association physical association false false false 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 1764932 n/a 1764936 unspecified role unspecified role prey prey n/a 1764938 unspecified role unspecified role prey prey n/a 1764937 unspecified role unspecified role bait bait n/a 1764939 unspecified role unspecified role prey prey physical association physical association false false false 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 1764932 n/a 1764941 unspecified role unspecified role bait bait n/a 1764942 unspecified role unspecified role prey prey n/a 1764943 unspecified role unspecified role prey prey n/a 1764939 unspecified role unspecified role prey prey n/a 1764940 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association false false false 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 1764932 n/a 1764944 unspecified role unspecified role bait bait n/a 1764945 unspecified role unspecified role prey prey n/a 1764940 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association false false false 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. gosr1-bet1-1 1764932 n/a 1764936 unspecified role unspecified role bait bait n/a 1764945 unspecified role unspecified role prey prey n/a 1764939 unspecified role unspecified role prey prey n/a 1764938 unspecified role unspecified role prey prey n/a 1764943 unspecified role unspecified role prey prey n/a 1764937 unspecified role unspecified role prey prey physical association physical association false false false Fig. 2a (lane 2), 2c gosr1-bet1-4 1764932 n/a 1764936 unspecified role unspecified role bait bait n/a 1764944 unspecified role unspecified role prey prey n/a 1764939 unspecified role unspecified role prey prey n/a 1764937 unspecified role unspecified role prey prey n/a 1764945 unspecified role unspecified role prey prey n/a 1764938 unspecified role unspecified role prey prey n/a 1764943 unspecified role unspecified role prey prey n/a 1764940 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association false false false 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 1764932 n/a 1764946 unspecified role unspecified role prey prey n/a 1764944 unspecified role unspecified role prey prey n/a 1764936 unspecified role unspecified role prey prey n/a 1764941 unspecified role unspecified role prey prey n/a 1764942 unspecified role unspecified role prey prey n/a 1764939 unspecified role unspecified role bait bait n/a 1764943 unspecified role unspecified role prey prey n/a 1764937 unspecified role unspecified role prey prey n/a 1764938 unspecified role unspecified role prey prey n/a 1764945 unspecified role unspecified role prey prey n/a 1764940 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association false false false When p115 is included, the SNARE bands are stronger. Vti1a bands are very faint. Fig. 2b (lanes 3-6), 2d stx5-bet1-2 1764932 n/a 1764936 unspecified role unspecified role prey prey n/a 1764946 unspecified role unspecified role prey prey n/a 1764945 unspecified role unspecified role prey prey n/a 1764942 unspecified role unspecified role prey prey n/a 1764939 unspecified role unspecified role bait bait n/a 1764937 unspecified role unspecified role prey prey n/a 1764938 unspecified role unspecified role prey prey n/a 1764943 unspecified role unspecified role prey prey n/a 1764941 unspecified role unspecified role prey prey physical association physical association false false false Fig. 2b (lane 2), 2d Vti1a bands are very faint. gosr1-bet1-3 1764932 n/a 1764936 unspecified role unspecified role bait bait n/a 1764938 unspecified role unspecified role prey prey n/a 1764937 unspecified role unspecified role prey prey n/a 1764943 unspecified role unspecified role prey prey n/a 1764939 unspecified role unspecified role prey prey physical association physical association false false false Fig. 2e (lane 2) stx5-bet1-5 1764932 n/a 1764936 unspecified role unspecified role prey prey n/a 1764941 unspecified role unspecified role prey prey n/a 1764942 unspecified role unspecified role prey prey n/a 1764937 unspecified role unspecified role prey prey n/a 1764943 unspecified role unspecified role prey prey n/a 1764939 unspecified role unspecified role bait bait n/a 1764938 unspecified role unspecified role prey prey physical association physical association false false false Fig. 2f (lane 2) stx5-uso1 1764933 n/a 1764940 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764939 unspecified role unspecified role bait bait cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range false 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 false ecoli Escherichia coli (strain K12) physical association physical association false false false 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 1764933 n/a 1764936 unspecified role unspecified role bait bait cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range false 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 false ecoli Escherichia coli (strain K12) n/a 1764940 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association false false false 1764933 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) stx5-uso1-1 1764933 n/a 1764939 unspecified role unspecified role bait bait cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range false 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 false ecoli Escherichia coli (strain K12) n/a 1764940 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 false rat-liver rattus norvegicus liver Rat liver liver 1765070 1765067 physical association physical association false false false 1764933 Fig. 6b (bottom left), 6c stx5-gosr1 1764933 n/a 1764936 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 false cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range false ecoli Escherichia coli (strain K12) n/a 1764940 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764939 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 false cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range false ecoli Escherichia coli (strain K12) physical association physical association false false false 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 1764933 n/a 1764936 unspecified role unspecified role bait bait cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range false 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 false ecoli Escherichia coli (strain K12) n/a 1764939 unspecified role unspecified role prey prey cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range false 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 false ecoli Escherichia coli (strain K12) n/a 1764940 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association false false false 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 1764933 n/a 1764936 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 false cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range false ecoli Escherichia coli (strain K12) n/a 1764939 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 false cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range false ecoli Escherichia coli (strain K12) 1765093 1765090 physical association physical association false false false 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 1764933 n/a 1764936 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 false cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range false ecoli Escherichia coli (strain K12) n/a 1764939 unspecified role unspecified role bait bait cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range false 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 false ecoli Escherichia coli (strain K12) 1765099 1765097 physical association physical association false false false 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 1764933 n/a 1764936 unspecified role unspecified role unspecified role unspecified role cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range false 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 false ecoli Escherichia coli (strain K12) n/a 1764939 unspecified role unspecified role unspecified role unspecified role cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range false 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 false ecoli Escherichia coli (strain K12) 1765106 1765103 physical association physical association false false false 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. uso1-bet1 1764934 n/a 1764944 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764936 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764940 unspecified role unspecified role bait bait biotin tag biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false rat-liver rattus norvegicus liver Rat liver liver n/a 1764937 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764943 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764942 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764945 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764941 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764938 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764939 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764947 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association false false false 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 1764934 n/a 1764936 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764942 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764941 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764939 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764943 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764938 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764937 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764947 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764940 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 false biotin tag biotin tag biotin 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) physical association physical association false false false 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 1764934 n/a 1764936 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764943 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764941 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764942 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764938 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764937 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764940 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 false biotin tag biotin tag biotin 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) physical association physical association false false false 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 1764934 n/a 1764944 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764936 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764945 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764941 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764942 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764939 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764937 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764940 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 false 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 false rat-liver rattus norvegicus liver Rat liver liver n/a 1764943 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764938 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764947 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association false false false 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 1764934 n/a 1764936 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764943 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764938 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764937 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764940 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 false tail domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false rat-liver rattus norvegicus liver Rat liver liver n/a 1764947 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764945 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764939 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764942 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver n/a 1764941 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association false false false 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-uso1-2 1764935 n/a 1764936 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 false cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind range range range range false ecoli Escherichia coli (strain K12) n/a 1764940 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 false rat-liver rattus norvegicus liver Rat liver liver 1765175 1765173 physical association physical association false false false Fig. 6b (top left)