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/ hui-2009-1 Synaptotagmin-mediated bending of the target membrane is a critical step in Ca(2+)-regulated fusion. 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 Cell (0092-8674) 2009 Only protein-protein interactions 3 imex curation hui-2009-2 Synaptotagmin-mediated bending of the target membrane is a critical step in Ca(2+)-regulated fusion. 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 Cell (0092-8674) 2009 Only protein-protein interactions 3 imex curation hui-2009-3 Synaptotagmin-mediated bending of the target membrane is a critical step in Ca(2+)-regulated fusion. 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 Cell (0092-8674) 2009 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 snp25_mouse Synaptosomal-associated protein 25 Synaptosomal-associated 25 kDa protein Super protein Snap25 Snap protein protein mouse Mus musculus Mouse MAEDADMRNELEEMQRRADQLADESLESTRRMLQLVEESKDAGIRTLVMLDEQGEQLERIEEGMDQINKDMKEAEKNLTDLGKFCGLCVCPCNKLKSSDAYKKAWGNNQDGVVASQPARVVDEREQMAISGGFIRRVTNDARENEMDENLEQVSGIIGNLRHMALDMGNEIDTQNRQIDRIMEKADSNKTRIDEANQRATKMLGSG syt1_rat Synaptotagmin-1 Syt1 Synaptotagmin I p65 protein protein rat Rattus norvegicus (Rat) Rat MVSASHPEALAAPVTTVATLVPHNATEPASPGEGKEDAFSKLKQKFMNELHKIPLPPWALIAIAIVAVLLVVTCCFCVCKKCLFKKKNKKKGKEKGGKNAINMKDVKDLGKTMKDQALKDDDAETGLTDGEEKEEPKEEEKLGKLQYSLDYDFQNNQLLVGIIQAAELPALDMGGTSDPYVKVFLLPDKKKKFETKVHRKTLNPVFNEQFTFKVPYSELGGKTLVMAVYDFDRFSKHDIIGEFKVPMNTVDFGHVTEEWRDLQSAEKEEQEKLGDICFSLRYVPTAGKLTVVILEAKNLKKMDVGGLSDPYVKIHLMQNGKRLKKKKTTIKKNTLNPYYNESFSFEVPFEQIQKVQVVVTVLDYDKIGKNDAIGKVFVGYNSTGAELRHWSDMLANPRRPIAQWHTLQVEEEVDAMLAVKK vamp2_mouse Vesicle-associated membrane protein 2 Vamp2 Syb2 Synaptobrevin-2 protein protein mouse Mus musculus Mouse MSATAATVPPAAPAGEGGPPAPPPNLTSNRRLQQTQAQVDEVVDIMRVNVDKVLERDQKLSELDDRADALQAGASQFETSAAKLKRKYWWKNLKMMIILGVICAIILIIIIVYFST syt1-snap25-1 Interaction between Syt1, Stx1a and Snap25 identirfied by coimmunoprecipitation 858812 n/a 858815 unspecified role unspecified role bait bait n/a 858816 unspecified role unspecified role prey prey n/a 858817 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 false 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 false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false cytoplasmic region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false 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 false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 false certain certain sequence position certain certain certain sequence position certain false 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 false certain certain sequence position certain certain certain sequence position certain false 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 false 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 false physical association physical association false false false Fig. 3A, 3B syt1-snap25-2 Interaction between Syt1, Stxa1 and Snap25 identirfied by coflotation 858813 n/a 858815 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 false n/a 858816 unspecified role unspecified role neutral component neutral component n/a 858817 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 false 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 false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false cytoplasmic region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false 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 false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig. 3C, 3D, 5B Increasing concentrations of Ca2+ stx1a-snap25-3 Interaction between Vamp2, Stxa1 and Snap25 identified by comigration 858814 n/a 858818 unspecified role unspecified role neutral component neutral component n/a 858815 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 false n/a 858816 unspecified role unspecified role neutral component neutral component physical association physical association false false false Supp. Fig. 9