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/ Synaptotagmin-mediated bending of the target membrane is a critical step in Ca(2+)-regulated fusion. Synaptotagmin-mediated bending of the target membrane is a critical step in Ca(2+)-regulated fusion. Cell (0092-8674) 2009 imex curation Hui E., Johnson CP., Yao J., Dunning FM., Chapman ER. Synapse - Interactions of proteins with an established role in the presynapse. Recombinant proteins (expression systems not given) were reconstituted into liposomes (25% Phosphatidylserine + 75% Phosphatidylcholine). Experimental details, and details of protein constructs and purification given in PMID: 14718921. SNAP25-binding activity of each syt protein was quantified by densitometry. chapman@physiology.wisc.edu 23-Dec-2009: Contacted by JYOTI. jyoti Only protein-protein interactions 3 in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip weight by comassie molecular weight estimation by coomasie staining weight by comassie Hui E., Johnson CP., Yao J., Dunning FM., Chapman ER. Synapse - Interactions of proteins with an established role in the presynapse. Recombinant proteins (expression systems not given) were reconstituted into liposomes (25% Phosphatidylserine + 75% Phosphatidylcholine). Experimental details, and details of protein constructs and purification given in PMID: 14718921. SNAP25-binding activity of each syt protein was quantified by densitometry. chapman@physiology.wisc.edu Accepted 2009-NOV-23 by JYOTI. 23-Dec-2009: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 3 imex curation Synaptotagmin-mediated bending of the target membrane is a critical step in Ca(2+)-regulated fusion. Synaptotagmin-mediated bending of the target membrane is a critical step in Ca(2+)-regulated fusion. Cell (0092-8674) 2009 imex curation Hui E., Johnson CP., Yao J., Dunning FM., Chapman ER. Synapse - Interactions of proteins with an established role in the presynapse. Recombinant proteins (expression systems not given) were reconstituted into liposomes (25% Phosphatidylserine + 75% Phosphatidylcholine). Experimental details, and details of protein constructs and purification given in PMID: 14718921. SNAP25-binding activity of each syt protein was quantified by densitometry. chapman@physiology.wisc.edu 23-Dec-2009: Contacted by JYOTI. jyoti Only protein-protein interactions 3 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 Hui E., Johnson CP., Yao J., Dunning FM., Chapman ER. Synapse - Interactions of proteins with an established role in the presynapse. Recombinant proteins (expression systems not given) were reconstituted into liposomes (25% Phosphatidylserine + 75% Phosphatidylcholine). Experimental details, and details of protein constructs and purification given in PMID: 14718921. SNAP25-binding activity of each syt protein was quantified by densitometry. chapman@physiology.wisc.edu Accepted 2009-NOV-23 by JYOTI. 23-Dec-2009: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 3 imex curation Synaptotagmin-mediated bending of the target membrane is a critical step in Ca(2+)-regulated fusion. Synaptotagmin-mediated bending of the target membrane is a critical step in Ca(2+)-regulated fusion. Cell (0092-8674) 2009 imex curation Hui E., Johnson CP., Yao J., Dunning FM., Chapman ER. Synapse - Interactions of proteins with an established role in the presynapse. Recombinant proteins (expression systems not given) were reconstituted into liposomes (25% Phosphatidylserine + 75% Phosphatidylcholine). Experimental details, and details of protein constructs and purification given in PMID: 14718921. SNAP25-binding activity of each syt protein was quantified by densitometry. chapman@physiology.wisc.edu 23-Dec-2009: Contacted by JYOTI. jyoti Only protein-protein interactions 3 in vitro In vitro affinity techniques affinity technology affinity techniques weight by comassie molecular weight estimation by coomasie staining weight by comassie Hui E., Johnson CP., Yao J., Dunning FM., Chapman ER. Synapse - Interactions of proteins with an established role in the presynapse. Recombinant proteins (expression systems not given) were reconstituted into liposomes (25% Phosphatidylserine + 75% Phosphatidylcholine). Experimental details, and details of protein constructs and purification given in PMID: 14718921. SNAP25-binding activity of each syt protein was quantified by densitometry. chapman@physiology.wisc.edu Accepted 2009-NOV-23 by JYOTI. 23-Dec-2009: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 3 imex curation stx1a_rat Syntaxin-1A Synaptotagmin-associated 35 kDa protein Neuron-specific antigen HPC-1 Stx1a Sap protein protein rat Rattus norvegicus (Rat) Rat MKDRTQELRTAKDSDDDDDVTVTVDRDRFMDEFFEQVEEIRGFIDKIAENVEEVKRKHSAILASPNPDEKTKEELEELMSDIKKTANKVRSKLKSIEQSIEQEEGLNRSSADLRIRKTQHSTLSRKFVEVMSEYNATQSDYRERCKGRIQRQLEITGRTTTSEELEDMLESGNPAIFASGIIMDSSISKQALSEIETRHSEIIKLENSIRELHDMFMDMAMLVESQGEMIDRIEYNVEHAVDYVERAVSDTKKAVKYQSKARRKKIMIIICCVILGIIIASTIGGIFG DIP protein P32851 original sequence version: 74 DIP protein P32851 original sequence version: 78 DIP protein P32851 original sequence version: 142 DIP protein P32851 original sequence version: 138 DIP protein P32851 original sequence version: 89 DIP protein P32851 original sequence version: 115 DIP protein P32851 original sequence version: 134 DIP protein P32851 original sequence version: 121 DIP protein P32851 original sequence version: 123 DIP protein P32851 original sequence version: 150 DIP protein P32851 original sequence version: 156 DIP protein P32851 original sequence version: 163 DIP protein P32851 original sequence version: 126 DIP protein P32851 original sequence version: 124 DIP protein P32851 original sequence version: 158 8F929F489E323CAA syt1_rat Synaptotagmin-1 Syt1 Synaptotagmin I p65 protein protein rat Rattus norvegicus (Rat) Rat MVSASHPEALAAPVTTVATLVPHNATEPASPGEGKEDAFSKLKQKFMNELHKIPLPPWALIAIAIVAVLLVVTCCFCVCKKCLFKKKNKKKGKEKGGKNAINMKDVKDLGKTMKDQALKDDDAETGLTDGEEKEEPKEEEKLGKLQYSLDYDFQNNQLLVGIIQAAELPALDMGGTSDPYVKVFLLPDKKKKFETKVHRKTLNPVFNEQFTFKVPYSELGGKTLVMAVYDFDRFSKHDIIGEFKVPMNTVDFGHVTEEWRDLQSAEKEEQEKLGDICFSLRYVPTAGKLTVVILEAKNLKKMDVGGLSDPYVKIHLMQNGKRLKKKKTTIKKNTLNPYYNESFSFEVPFEQIQKVQVVVTVLDYDKIGKNDAIGKVFVGYNSTGAELRHWSDMLANPRRPIAQWHTLQVEEEVDAMLAVKK DIP protein P21707 original sequence version: 71 DIP protein P21707 original sequence version: 76 DIP protein P21707 original sequence version: 141 DIP protein P21707 original sequence version: 112 DIP protein P21707 original sequence version: 145 DIP protein P21707 original sequence version: 160 06CE28F04C97A722 vamp2_mouse Vesicle-associated membrane protein 2 Vamp2 Syb2 Synaptobrevin-2 protein protein mouse Mus musculus Mouse MSATAATVPPAAPAGEGGPPAPPPNLTSNRRLQQTQAQVDEVVDIMRVNVDKVLERDQKLSELDDRADALQAGASQFETSAAKLKRKYWWKNLKMMIILGVICAIILIIIIVYFST DIP protein P63044 original sequence version: 28 DIP protein P63044 original sequence version: 32 DIP protein P63044 original sequence version: 98 DIP protein P63044 original sequence version: 70 DIP protein P63044 original sequence version: 100 DIP protein P63044 original sequence version: 116 4A0D0D56B5409D0A snp25_mouse Synaptosomal-associated protein 25 Synaptosomal-associated 25 kDa protein Super protein Snap25 Snap protein protein mouse Mus musculus Mouse MAEDADMRNELEEMQRRADQLADESLESTRRMLQLVEESKDAGIRTLVMLDEQGEQLERIEEGMDQINKDMKEAEKNLTDLGKFCGLCVCPCNKLKSSDAYKKAWGNNQDGVVASQPARVVDEREQMAISGGFIRRVTNDARENEMDENLEQVSGIIGNLRHMALDMGNEIDTQNRQIDRIMEKADSNKTRIDEANQRATKMLGSG DIP protein P60879 original sequence version: 38 DIP protein P60879-1 original sequence version: 41 DIP protein P60879-1 original sequence version: 109 DIP protein P60879-1 original sequence version: 81 DIP protein P60879 original sequence version: 111 FBED2B082A4CB6A6 syt1-snap25-1 1 4 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey region 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 undefined mutation M173W, F234W, V304W, I367W mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:34:00 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'c2a2w-c2b2w' undefined mutation M173A, F234A, V304A, I367A mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:33:55 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'c2a2a-c2b2a' cytoplasmic region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain undefined mutation M173W, F231W, F234W, V304W, I364W, I367W mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:34:02 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'c2a3w-c2b3w' undefined mutation D230N, D232N mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:33:54 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'c2aclm-c2b' undefined mutation D363N, D365N mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:33:53 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'c2a-c2bclm' undefined mutation region 272-461->region 142-263 mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing substitut analysis substitution analysis substitut analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Wed Jul 06 16:33:22 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'c2a-c2a' [EBI-2462732]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 461-461, sequence length 421 [EBI-2462732]272-461 undefined mutation region 96-263->273-461 mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing substitut analysis substitution analysis substitut analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:33:22 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'c2b-c2b' physical association physical association Fig. 3A, 3B Coimmunoprecipitation of syt fragments with t-SNARE heterodimers was carried out using an anti-syntaxin (syx) antibody. syt1-snap25-2 3 4 unspecified role unspecified role neutral component neutral component region 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 7 unspecified role unspecified role neutral component neutral component 5 unspecified role unspecified role neutral component neutral component region 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 undefined mutation M173W, F234W, V304W, I367W mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:33:12 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'c2a2w-c2b2w' undefined mutation M173A, F234A, V304A, I367A mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:33:07 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'c2a2a-c2b2a' cytoplasmic region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain undefined mutation M173W, F231W, F234W, V304W, I364W, I367W mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:33:32 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'c2a3w-c2b3w' physical association physical association Fig. 3C, 3D, 5B Trp/Ala mutations within the membrane-penetration loops of syt strongly affect syt's ability to engage t-SNAREs in response to Ca2+, as assessed using co-flotation assays with t-SNARE-bearing, PS-free vesicles. Increasing concentrations of Ca2+ stx1a-snap25-3 2 6 unspecified role unspecified role neutral component neutral component 4 unspecified role unspecified role neutral component neutral component region 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 7 unspecified role unspecified role neutral component neutral component physical association physical association Supp. Fig. 9 For both t-SNARE GUVs and SUVs, increasing [cd-syb] efficiently shifted the t-SNARE hetero-dimer into SDS-resistant ternary SNARE complexes.