UniProt UniProt http://www.uniprot.org Syntaxin 17 promotes lipid droplet formation by regulating the distribution of acyl-CoA synthetase 3. Syntaxin 17 promotes lipid droplet formation by regulating the distribution of acyl-CoA synthetase 3. Journal of lipid research 2018 imex curation Kimura H., Arasaki K., Ohsaki Y., Fujimoto T., Ohtomo T., Yamada J., Tagaya M. Source pf LPCAT1 could not be traced tagaya@toyaku.ac.jp 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Kimura H., Arasaki K., Ohsaki Y., Fujimoto T., Ohtomo T., Yamada J., Tagaya M. Source pf LPCAT1 could not be traced tagaya@toyaku.ac.jp Accepted 2020-FEB-25 AT 10:16 GMT AT 10:16 GMT by PPORRAS Journal of lipid research 2018 Only protein-protein interactions imex curation imex curation Syntaxin 17 promotes lipid droplet formation by regulating the distribution of acyl-CoA synthetase 3. Syntaxin 17 promotes lipid droplet formation by regulating the distribution of acyl-CoA synthetase 3. Journal of lipid research 2018 imex curation Kimura H., Arasaki K., Ohsaki Y., Fujimoto T., Ohtomo T., Yamada J., Tagaya M. Source pf LPCAT1 could not be traced tagaya@toyaku.ac.jp 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Kimura H., Arasaki K., Ohsaki Y., Fujimoto T., Ohtomo T., Yamada J., Tagaya M. Source pf LPCAT1 could not be traced tagaya@toyaku.ac.jp Accepted 2020-FEB-25 AT 10:16 GMT AT 10:16 GMT by PPORRAS Journal of lipid research 2018 Only protein-protein interactions imex curation imex curation Syntaxin 17 promotes lipid droplet formation by regulating the distribution of acyl-CoA synthetase 3. Syntaxin 17 promotes lipid droplet formation by regulating the distribution of acyl-CoA synthetase 3. Journal of lipid research 2018 imex curation Kimura H., Arasaki K., Ohsaki Y., Fujimoto T., Ohtomo T., Yamada J., Tagaya M. Source pf LPCAT1 could not be traced tagaya@toyaku.ac.jp 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Kimura H., Arasaki K., Ohsaki Y., Fujimoto T., Ohtomo T., Yamada J., Tagaya M. Source pf LPCAT1 could not be traced tagaya@toyaku.ac.jp Accepted 2020-FEB-25 AT 10:16 GMT AT 10:16 GMT by PPORRAS Journal of lipid research 2018 Only protein-protein interactions imex curation imex curation snp23_human Synaptosomal-associated protein 23 Vesicle-membrane fusion protein SNAP-23 SNAP23 protein protein human Homo sapiens Human MDNLSSEEIQQRAHQITDESLESTRRILGLAIESQDAGIKTITMLDEQKEQLNRIEEGLDQINKDMRETEKTLTELNKCCGLCVCPCNRTKNFESGKAYKTTWGDGGENSPCNVVSKQPGPVTNGQLQQPTTGAASGGYIKRITNDAREDEMEENLTQVGSILGNLKDMALNIGNEIDAQNPQIKRITDKADTNRDRIDIANARAKKLIDS AC378E9786C3A239 stx17_human Syntaxin-17 STX17 protein protein human Homo sapiens Human MSEDEEKVKLRRLEPAIQKFIKIVIPTDLERLRKHQINIEKYQRCRIWDKLHEEHINAGRTVQQLRSNIREIEKLCLKVRKDDLVLLKRMIDPVKEEASAATAEFLQLHLESVEELKKQFNDEETLLQPPLTRSMTVGGAFHTTEAEASSQSLTQIYALPEIPQDQNAAESWETLEADLIELSQLVTDFSLLVNSQQEKIDSIADHVNSAAVNVEEGTKNLGKAAKYKLAALPVAGALIGGMVGGPIGLLAGFKVAGIAAALGGGVLGFTGGKLIQRKKQKMMEKLTSSCPDLPSQTDKKCS DIP protein P56962 original sequence version: 116 DIP protein P56962 original sequence version: 86 51AD5427EFBAC541 acsl3_human Fatty acid CoA ligase Acsl3 Long-chain acyl-CoA synthetase 3 ACSL3 ACS3 FACL3 LACS3 Arachidonate--CoA ligase Long-chain-fatty-acid--CoA ligase 3 Medium-chain acyl-CoA ligase Acsl3 protein protein human Homo sapiens Human MNNHVSSKPSTMKLKHTINPILLYFIHFLISLYTILTYIPFYFFSESRQEKSNRIKAKPVNSKPDSAYRSVNSLDGLASVLYPGCDTLDKVFTYAKNKFKNKRLLGTREVLNEEDEVQPNGKIFKKVILGQYNWLSYEDVFVRAFNFGNGLQMLGQKPKTNIAIFCETRAEWMIAAQACFMYNFQLVTLYATLGGPAIVHALNETEVTNIITSKELLQTKLKDIVSLVPRLRHIITVDGKPPTWSEFPKGIIVHTMAAVEALGAKASMENQPHSKPLPSDIAVIMYTSGSTGLPKGVMISHSNIIAGITGMAERIPELGEEDVYIGYLPLAHVLELSAELVCLSHGCRIGYSSPQTLADQSSKIKKGSKGDTSMLKPTLMAAVPEIMDRIYKNVMNKVSEMSSFQRNLFILAYNYKMEQISKGRNTPLCDSFVFRKVRSLLGGNIRLLLCGGAPLSATTQRFMNICFCCPVGQGYGLTESAGAGTISEVWDYNTGRVGAPLVCCEIKLKNWEEGGYFNTDKPHPRGEILIGGQSVTMGYYKNEAKTKADFFEDENGQRWLCTGDIGEFEPDGCLKIIDRKKDLVKLQAGEYVSLGKVEAALKNLPLVDNICAYANSYHSYVIGFVVPNQKELTELARKKGLKGTWEELCNSCEMENEVLKVLSEAAISASLEKFEIPVKIRLSPEPWTPETGLVTDAFKLKRKELKTHYQADIERMYGRK AAC4B0B4543EC8DD acsl3-stx17-1 1 6 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed P56962:p.Lys254Cys mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain K C Tue Jun 30 16:14:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys254Cys' region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Fig. 6A Significant amounts of endogenous ACSL3 were coprecipitated with FLAG-Stx17 wild-type and the K254C mutant, but not with the ΔSNARE mutant (Fig. 6A) stx17-acsl3-1 2 6 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level 5 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed P56962:p.Lys254Cys mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain K C Tue Jun 30 17:52:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys254Cys' region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range proximity proximity close-contact colocalization neighbourhood interaction Fig. 6B, 6C, 6D, Similar results were obtained for PLA (Fig. 6B), suggesting that Stx17 interacts with ACSL3 via its SNARE domain. Triacsin C (CHEBI:144441) Diacylglycerol O-acyltransferase inhibitors (CHEBI:64919) Oleic acid (CHEBI:16196) stx17-acsl3-2 3 6 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain n-terminal egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Fig. 6E Binding experiments revealed that both the TMD and GATE domains of ACSL3 are required for the interaction with Stx17 (Fig. 6E) stx17-acsl3-3 2 6 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain n-terminal egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 5 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range proximity proximity close-contact colocalization neighbourhood interaction Fig. 6F Oleic acid (CHEBI:16196) stx17-snap23-1 1 4 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed P56962:p.Lys254Cys mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain K C Tue Jun 30 16:39:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys254Cys' region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Fig. 8A We determined the region of Stx17 responsible for the interaction with SNAP23 by immunoprecipitation. As expected, the interaction of Stx17 with SNAP23 was abolished on deletion of the SNARE domain of Stx17 (Fig. 8A, lane 8). Interestingly, the interaction was drastically reduced on mutation at Lys254 located in the middle of the CHD, even though the resultant mutant retained the SNARE domain (Fig. 8A, lane 7). stx17-snap23-2 2 4 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level 5 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level proximity proximity close-contact colocalization neighbourhood interaction Fig. 8E Oleic acid (CHEBI:16196) acsl3-stx17-2 2 6 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level 5 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level proximity proximity close-contact colocalization neighbourhood interaction Fig. 8B, 8C, 8E Oleic acid (CHEBI:16196) When SNAP23 was knocked down, the number of PLA dots representing the proximity between Stx17 and ACSL3 at the endogenous level was significantly increased, whereas ectopic expression of SNAP23 reduced the number of the PLA dots.