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/ kioumourtzoglou-2014-1 Insulin stimulates syntaxin4 SNARE complex assembly via a novel regulatory mechanism. mouse-3t3_l1 Mus musculus embryonic cells Fibroblast-like. Adipose like when confluent 3t3_l1 Mouse embryo cells Fibroblast-like. Adipose like when confluent pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Kioumourtzoglou D., Gould G.W., Bryant N.J. Diabetes - Interactions investigated in the context of Diabetes Results from Figure 4 and Supp Fig 4 have not been included as both Munc18c and SNAP23 have His-tags, so the individual constituents of the complex cannot be identified; neither is it possible to identify the presence of individual proteins from the Coommasie blue stained gel, as the bands coalesce. Results from Supp. Fig 3-2 have not been added, as only some components of the complex are detected. The authors have stated that:- [1] Sx4 (Syntaxin4) is of RATTINE origin. [2] SNAP23 is of MURINE origin and the protein has 210 aa's; the statement in table 1 that SNAP23 has 211 aa's is a typo. [3] All Munc18c species used were MURINE. The authors also confirm that all Syntaxin 4 cytoplasmic domain constructs are made from amino acids 1-273, as stated in Table 1, not amino acids 1-211 as given in the text above Figure 2. gwyn.gould@glasgow.ac.uk, nia.bryant@york.ac.uk Mol. Cell. Biol. (0270-7306) 2014 Only protein-protein interactions imex curation Affinomics - Interactions curated for the Affinomics consortium. kioumourtzoglou-2014-2 Insulin stimulates syntaxin4 SNARE complex assembly via a novel regulatory mechanism. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Kioumourtzoglou D., Gould G.W., Bryant N.J. Diabetes - Interactions investigated in the context of Diabetes Results from Figure 4 and Supp Fig 4 have not been included as both Munc18c and SNAP23 have His-tags, so the individual constituents of the complex cannot be identified; neither is it possible to identify the presence of individual proteins from the Coommasie blue stained gel, as the bands coalesce. Results from Supp. Fig 3-2 have not been added, as only some components of the complex are detected. The authors have stated that:- [1] Sx4 (Syntaxin4) is of RATTINE origin. [2] SNAP23 is of MURINE origin and the protein has 210 aa's; the statement in table 1 that SNAP23 has 211 aa's is a typo. [3] All Munc18c species used were MURINE. The authors also confirm that all Syntaxin 4 cytoplasmic domain constructs are made from amino acids 1-273, as stated in Table 1, not amino acids 1-211 as given in the text above Figure 2. gwyn.gould@glasgow.ac.uk, nia.bryant@york.ac.uk Mol. Cell. Biol. (0270-7306) 2014 Only protein-protein interactions imex curation Affinomics - Interactions curated for the Affinomics consortium. kioumourtzoglou-2014-4 Insulin stimulates syntaxin4 SNARE complex assembly via a novel regulatory mechanism. in vitro In vitro filter binding filter binding Filter overlay assay Filter overlay assay western blot western blot Immuno blot Immuno blot Kioumourtzoglou D., Gould G.W., Bryant N.J. Diabetes - Interactions investigated in the context of Diabetes Results from Figure 4 and Supp Fig 4 have not been included as both Munc18c and SNAP23 have His-tags, so the individual constituents of the complex cannot be identified; neither is it possible to identify the presence of individual proteins from the Coommasie blue stained gel, as the bands coalesce. Results from Supp. Fig 3-2 have not been added, as only some components of the complex are detected. The authors have stated that:- [1] Sx4 (Syntaxin4) is of RATTINE origin. [2] SNAP23 is of MURINE origin and the protein has 210 aa's; the statement in table 1 that SNAP23 has 211 aa's is a typo. [3] All Munc18c species used were MURINE. The authors also confirm that all Syntaxin 4 cytoplasmic domain constructs are made from amino acids 1-273, as stated in Table 1, not amino acids 1-211 as given in the text above Figure 2. gwyn.gould@glasgow.ac.uk, nia.bryant@york.ac.uk Mol. Cell. Biol. (0270-7306) 2014 Only protein-protein interactions imex curation Affinomics - Interactions curated for the Affinomics consortium. kioumourtzoglou-2014-3 Insulin stimulates syntaxin4 SNARE complex assembly via a novel regulatory mechanism. in vitro In vitro comigration in sds comigration in sds page comigration in sds weight by comassie molecular weight estimation by coomasie staining weight by comassie Kioumourtzoglou D., Gould G.W., Bryant N.J. Diabetes - Interactions investigated in the context of Diabetes Results from Figure 4 and Supp Fig 4 have not been included as both Munc18c and SNAP23 have His-tags, so the individual constituents of the complex cannot be identified; neither is it possible to identify the presence of individual proteins from the Coommasie blue stained gel, as the bands coalesce. Results from Supp. Fig 3-2 have not been added, as only some components of the complex are detected. The authors have stated that:- [1] Sx4 (Syntaxin4) is of RATTINE origin. [2] SNAP23 is of MURINE origin and the protein has 210 aa's; the statement in table 1 that SNAP23 has 211 aa's is a typo. [3] All Munc18c species used were MURINE. The authors also confirm that all Syntaxin 4 cytoplasmic domain constructs are made from amino acids 1-273, as stated in Table 1, not amino acids 1-211 as given in the text above Figure 2. gwyn.gould@glasgow.ac.uk, nia.bryant@york.ac.uk Mol. Cell. Biol. (0270-7306) 2014 Only protein-protein interactions imex curation Affinomics - Interactions curated for the Affinomics consortium. stxb3_mouse Syntaxin-binding protein 3 Stxbp3 Stxbp3a Unc18c Protein unc-18 homolog 3 Protein unc-18 homolog C Mammalian homolog of Unc-18c MUNC-18-3 protein protein mouse Mus musculus Mouse MAPPVSERGLKSVVWRKIKTAVFDDCRKEGEWKIMLLDEFTTKLLSSCCKMTDLLEEGITVIENIYKNREPVRQMKALYFISPTPKSVDCFLRDFGSKSEKKYKAAYIYFTDFCPDSLFNKIKASCSKSIRRCKEINISFIPQESQVYTLDVPDAFYYCYSPDPSNASRKEVVMEAMAEQIVTVCATLDENPGVRYKSKPLDNASKLAQLVEKKLEDYYKIDEKGLIKGKTQSQLLIIDRGFDPVSTVLHELTFQAMAYDLLPIENDTYKYKTDGKEKEAVLEEDDDLWVRVRHRHIAVVLEEIPKLMKEISSTKKATEGKTSLSALTQLMKKMPHFRKQISKQVVHLNLAEDCMNKFKLNIEKLCKTEQDLALGTDAEGQRVKDSMLVLLPVLLNKNHDNCDKIRAVLLYIFGINGTTEENLDRLIHNVKIEDDSDMIRNWSHLGVPIVPPSQQAKPLRKDRSAEETFQLSRWTPFIKDIMEDAIDNRLDSKEWPYCSRCPAVWNGSGAVSARQKPRTNYLELDRKNGSRLIIFVIGGITYSEMRCAYEVSQAHKSCEVIIGSTHILTPRKLLDDIKMLNKSKDKVSFKDE mint stx4_mouse Syntaxin-4 Stx4 Stx4a protein protein mouse Mus musculus Mouse MRDRTHELRQGDNISDDEDEVRVALVVHSGAARLGSPDDEFFQKVQTIRQTMAKLESKVRELEKQQVTILATPLPEESMKQGLQNLREEIKQLGREVRAQLKAIEPQKEEADENYNSVNTRMKKTQHGVLSQQFVELINKCNSMQSEYREKNVERIRRQLKITNAGMVSDEELEQMLDSGQSEVFVSNILKDTQVTRQALNEISARHSEIQQLERSIRELHEIFTFLATEVEMQGEMINRIEKNILSSADYVERGQEHVKIALENQKKARKKKVMIAICVSVTVLILAVIIGITITVG vamp2_mouse Vesicle-associated membrane protein 2 Vamp2 Syb2 Synaptobrevin-2 protein protein mouse Mus musculus Mouse MSATAATVPPAAPAGEGGPPAPPPNLTSNRRLQQTQAQVDEVVDIMRVNVDKVLERDQKLSELDDRADALQAGASQFETSAAKLKRKYWWKNLKMMIILGVICAIILIIIIVYFST snp23_mouse Synaptosomal-associated protein 23 Snap23 Sndt Vesicle-membrane fusion protein SNAP-23 Syndet protein protein mouse Mus musculus Mouse MDNLSPEEVQLRAHQVTDESLESTRRILGLAIESQDAGIKTITMLDEQGEQLNRIEEGMDQINKDMREAEKTLTELNKCCGLCICPCNRTKNFESGKNYKATWGDGGDNSPSNVVSKQPSRITNGQPQQTTGAASGGYIKRITNDAREDEMEENLTQVGSILGNLKNMALDMGNEIDAQNQQIQKITEKADTNKNRIDIANTRAKKLIDS stx4_rat Syntaxin-4 Stx4 Stx4a protein protein rat Rattus norvegicus (Rat) Rat MRDRTHELRQGDNISDDEDEVRVALVVHSGAARLSSPDDEFFQKVQTIRQTMAKLESKVRELEKQQVTILATPLPEESMKQGLQNLREEIKQLGREVRAQLKAIEPQKEEADENYNSVNTRMKKTQHGVLSQQFVELINKCNSMQSEYREKNVERIRRQLKITNAGMVSDEELEQMLDSGQSEVFVSNILKDTQVTRQALNEISARHSEIQQLERSIRELHEIFTFLATEVEMQGEMINRIEKNILSSADYVERGQEHVKIALENQKKARKKKVMIAICVSVTVLILAVIIGITITVG vamp2_rat Vesicle-associated membrane protein 2 Vamp2 Syb2 Synaptobrevin-2 protein protein rat Rattus norvegicus (Rat) Rat MSATAATVPPAAPAGEGGPPAPPPNLTSNRRLQQTQAQVDEVVDIMRVNVDKVLERDQKLSELDDRADALQAGASQFETSAAKLKRKYWWKNLKMMIILGVICAIILIIIIVYFST stx4-stxbp3 1573170 n/a 1573174 unspecified role unspecified role neutral component neutral component n/a 1573175 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 1 stx4-stxbp3-1 1573170 n/a 1573174 unspecified role unspecified role neutral component neutral component n/a 1573176 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 1 stxbp3-vamp2 1573170 n/a 1573174 unspecified role unspecified role neutral component neutral component n/a 1573177 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 1 Insulin snap23-stxbp3 1573170 n/a 1573175 unspecified role unspecified role neutral component neutral component n/a 1573177 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 1 snap23-stx4-7 1573170 n/a 1573176 unspecified role unspecified role neutral component neutral component n/a 1573175 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 1 stx4-vamp2 1573170 n/a 1573176 unspecified role unspecified role neutral component neutral component n/a 1573177 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false 1 Insulin vamp2-stx4 1573171 n/a 1573178 unspecified role unspecified role prey prey cytosolic_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1573179 unspecified role unspecified role bait bait cytosolic_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c_terminus protein a tag protein a tag protein A protein A c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) 1573202 1573200 direct interaction direct interaction false false false 2A stx4-vamp2-1 1573171 n/a 1573178 unspecified role unspecified role bait bait snare_coiled-coil_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false Q08850:p.Leu173_Glu174delinsAlaAla mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1573179 unspecified role unspecified role prey prey c_terminus protein a tag protein a tag protein A protein A c-term range C-terminal range c-term range c-term range C-terminal range c-term range false cytosolic_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 1573211 1573206 direct interaction direct interaction false false false 2B/C/D/E vamp2-stx4-1 1573171 n/a 1573174 unspecified role unspecified role bait bait n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1573178 unspecified role unspecified role prey prey cytosolic_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 1573217 1573215 direct interaction direct interaction false false false Supp. Fig 1-2 C stxbp3-stx4 1573171 n/a 1573177 unspecified role unspecified role bait bait n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst 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) n/a 1573178 unspecified role unspecified role prey prey cytosolic_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Supp. Fig 1-2 D snap23-stx4-9 1573171 n/a 1573177 unspecified role unspecified role bait bait n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-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) n/a 1573178 unspecified role unspecified role prey prey cytosolic_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Supp. Fig 1-2 E stx4-snap23-2 1573171 n/a 1573178 unspecified role unspecified role bait bait cytosolic_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1573177 unspecified role unspecified role prey prey n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1573179 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey c_terminus protein a tag protein a tag protein A protein A c-term range C-terminal range c-term range c-term range C-terminal range c-term range false cytosolic_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 1573236 1573230 association association false false false Supp Fig 3-1 stx4-vamp2-2 1573172 n/a 1573178 unspecified role unspecified role bait bait cytosolic_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1573179 unspecified role unspecified role prey prey c_terminus protein a tag protein a tag protein A protein A c-term range C-terminal range c-term range c-term range C-terminal range c-term range false cytosolic_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 1573242 1573239 direct interaction direct interaction false false false Supp Fig 2A stx4-vamp2-3 1573172 n/a 1573179 unspecified role unspecified role prey prey v-snare coiled-coil homology_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1573178 unspecified role unspecified role bait bait snare_coiled-coil_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 1573247 1573245 direct interaction direct interaction false false false Supp Fig 2B snap23-stx4-8 1573173 n/a 1573177 unspecified role unspecified role neutral component neutral component n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-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) n/a 1573178 unspecified role unspecified role neutral component neutral component cytosolic_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1573179 western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false cytosolic_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false 3