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/ Phosphorylation of SNAP-23 by IkappaB kinase 2 regulates mast cell degranulation. Phosphorylation of SNAP-23 by IkappaB kinase 2 regulates mast cell degranulation. Cell (0092-8674) 2008 imex curation Suzuki K., Verma IM. Synapse - Interactions of proteins with an established role in the presynapse. Samples pre-fractionated by sucrose density gradient centrifugation verma@salk.edu 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 6 rat-rbl_2h3 Rat basophilic leukemia cell line rbl_2h3 Rat basophilic leukemia cell line anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Suzuki K., Verma IM. Synapse - Interactions of proteins with an established role in the presynapse. Samples pre-fractionated by sucrose density gradient centrifugation verma@salk.edu Accepted 2008-OCT-23 by DAVET. 16-Oct-2008: Contacted by JYOTI. Fcepsilon R1 (P30273) stimulation Cell (0092-8674) 2008 davet Only protein-protein interactions 6 imex curation Phosphorylation of SNAP-23 by IkappaB kinase 2 regulates mast cell degranulation. Phosphorylation of SNAP-23 by IkappaB kinase 2 regulates mast cell degranulation. Cell (0092-8674) 2008 imex curation Suzuki K., Verma IM. Synapse - Interactions of proteins with an established role in the presynapse. Samples pre-fractionated by sucrose density gradient centrifugation verma@salk.edu 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 6 rat-rbl_2h3 Rat basophilic leukemia cell line rbl_2h3 Rat basophilic leukemia cell line density sedimentation cosedimentation through density gradient density sedimentation western blot western blot Immuno blot Immuno blot Suzuki K., Verma IM. Synapse - Interactions of proteins with an established role in the presynapse. Samples pre-fractionated by sucrose density gradient centrifugation verma@salk.edu Accepted 2008-OCT-23 by DAVET. 16-Oct-2008: Contacted by JYOTI. Fcepsilon R1 (P30273) stimulation Cell (0092-8674) 2008 davet Only protein-protein interactions 6 imex curation Phosphorylation of SNAP-23 by IkappaB kinase 2 regulates mast cell degranulation. Phosphorylation of SNAP-23 by IkappaB kinase 2 regulates mast cell degranulation. Cell (0092-8674) 2008 imex curation Suzuki K., Verma IM. Synapse - Interactions of proteins with an established role in the presynapse. Samples pre-fractionated by sucrose density gradient centrifugation verma@salk.edu 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 6 mouse-mast_cell Mouse mast cell mast_cell Mast cell anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Suzuki K., Verma IM. Synapse - Interactions of proteins with an established role in the presynapse. Samples pre-fractionated by sucrose density gradient centrifugation verma@salk.edu Accepted 2008-OCT-23 by DAVET. 16-Oct-2008: Contacted by JYOTI. Fcepsilon R1 (P30273) stimulation Cell (0092-8674) 2008 davet Only protein-protein interactions 6 imex curation Phosphorylation of SNAP-23 by IkappaB kinase 2 regulates mast cell degranulation. Phosphorylation of SNAP-23 by IkappaB kinase 2 regulates mast cell degranulation. Cell (0092-8674) 2008 imex curation Suzuki K., Verma IM. Synapse - Interactions of proteins with an established role in the presynapse. Samples pre-fractionated by sucrose density gradient centrifugation verma@salk.edu 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 6 in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Suzuki K., Verma IM. Synapse - Interactions of proteins with an established role in the presynapse. Samples pre-fractionated by sucrose density gradient centrifugation verma@salk.edu Accepted 2008-OCT-23 by DAVET. 16-Oct-2008: Contacted by JYOTI. Fcepsilon R1 (P30273) stimulation Cell (0092-8674) 2008 davet Only protein-protein interactions 6 imex curation Phosphorylation of SNAP-23 by IkappaB kinase 2 regulates mast cell degranulation. Phosphorylation of SNAP-23 by IkappaB kinase 2 regulates mast cell degranulation. Cell (0092-8674) 2008 imex curation Suzuki K., Verma IM. Synapse - Interactions of proteins with an established role in the presynapse. Samples pre-fractionated by sucrose density gradient centrifugation verma@salk.edu 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 6 human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Suzuki K., Verma IM. Synapse - Interactions of proteins with an established role in the presynapse. Samples pre-fractionated by sucrose density gradient centrifugation Source of IKK2 confirmed by author. verma@salk.edu Accepted 2008-OCT-23 by DAVET. 16-Oct-2008: Contacted by JYOTI. Fcepsilon R1 (P30273) stimulation Cell (0092-8674) 2008 davet Only protein-protein interactions 6 imex curation stx4_mouse Syntaxin-4 Stx4 Stx4a protein protein mouse Mus musculus Mouse MRDRTHELRQGDNISDDEDEVRVALVVHSGAARLGSPDDEFFQKVQTIRQTMAKLESKVRELEKQQVTILATPLPEESMKQGLQNLREEIKQLGREVRAQLKAIEPQKEEADENYNSVNTRMKKTQHGVLSQQFVELINKCNSMQSEYREKNVERIRRQLKITNAGMVSDEELEQMLDSGQSEVFVSNILKDTQVTRQALNEISARHSEIQQLERSIRELHEIFTFLATEVEMQGEMINRIEKNILSSADYVERGQEHVKIALENQKKARKKKVMIAICVSVTVLILAVIIGITITVG DIP protein P70452 original sequence version: 120 DIP protein P70452 original sequence version: 108 FCD1477E1126CEC1 vamp8_mouse Vesicle-associated membrane protein 8 Vamp8 Endobrevin protein protein mouse Mus musculus Mouse MEEASGSAGNDRVRNLQSEVEGVKNIMTQNVERILSRGENLDHLRNKTEDLEATSEHFKTTSQKVARKFWWKNVKMIVIICVIVLIIVILIILFATGTIPT 0A2FDB6B69E06C3E stx4_rat Syntaxin-4 Stx4 Stx4a protein protein rat Rattus norvegicus (Rat) Rat MRDRTHELRQGDNISDDEDEVRVALVVHSGAARLSSPDDEFFQKVQTIRQTMAKLESKVRELEKQQVTILATPLPEESMKQGLQNLREEIKQLGREVRAQLKAIEPQKEEADENYNSVNTRMKKTQHGVLSQQFVELINKCNSMQSEYREKNVERIRRQLKITNAGMVSDEELEQMLDSGQSEVFVSNILKDTQVTRQALNEISARHSEIQQLERSIRELHEIFTFLATEVEMQGEMINRIEKNILSSADYVERGQEHVKIALENQKKARKKKVMIAICVSVTVLILAVIIGITITVG E98547758FE6CEC6 ikkb_rat Inhibitor of nuclear factor kappa-B kinase subunit beta Ikbkb Ikkb I-kappa-B kinase 2 Nuclear factor NF-kappa-B inhibitor kinase beta Serine/threonine protein kinase IKBKB protein protein rat Rattus norvegicus (Rat) Rat MSWSPSLPTQTCGAWEMKERLGTGGFGNVIRWHNQVTGEQIAIKQCRQELSPKNRDRWCLEIQIMRRLNHPNVVAARDVPEGMQNLAPNDLPLLAMEYCQGGDLRRYLNQFENCCGLREGAILTLLSDIASALRYLHENRIIHRDLKPENIVLQQGEKRLIHKIIDLGYAKELDQGSLCTSFVGTLQYLAPELLEQQKYTVTVDYWSFGTLAFECITGFRPFLPNWQPVQWHSKVRQKSEVDIVVSEDLNGTVKFSSSSPFPNNLNSVLAERLEKWLQLMLTWQPRQRGVDPQYGPNGCFRALDDILNLKLVHILNMVTGTIHTYPVMEDESLQSLKTRIREDTGILETDQELLQEAGLVLLPDKPATQCISDSKTNEGLTLDMDLVFLFDNSKMSYETQITPRPQPESVSCVLQEPKRNLSFFQMRKVWGQVWHSIQTLKEDCNRLQQGQRAAMMNLLRNNSCLSKMKNAMASTAQQLKAKLDFFKTSIQIDLEKYREQTEFGITSDKLLLAWREMEQAVEQCGRENDVKVLVERMMALQTDIVDLQRSPMGRKQGGTLDDLEEQARELYRRLREKPRDQRTEGDSQDMVRLLLQAIQSFEKKVRVIYSQLSKTVVCKQKALELLPKVEEVVRLMNEDEKTVVRLQEKRQKELWNLLKIACSKVRGPVSGSPDSMNVSRLSHPGHLMSQPSSACDSLPDSDKKSEELVAEAHALCSRLESALQDTVKQQDRSFTTLDWSWLQMEDEERCGLEQACD 3AFFE46A7DF91F9C vamp2_rat Vesicle-associated membrane protein 2 Vamp2 Syb2 Synaptobrevin-2 protein protein rat Rattus norvegicus (Rat) Rat MSATAATVPPAAPAGEGGPPAPPPNLTSNRRLQQTQAQVDEVVDIMRVNVDKVLERDQKLSELDDRADALQAGASQFETSAAKLKRKYWWKNLKMMIILGVICAIILIIIIVYFST DIP protein P63045 original sequence version: 95 DIP protein P63045 original sequence version: 90 DIP protein P63045 original sequence version: 64 DIP protein P63045 original sequence version: 62 DIP protein P63045 original sequence version: 85 DIP protein P63045 original sequence version: 104 DIP protein P63045 original sequence version: 77 DIP protein P63045 original sequence version: 75 DIP protein P63045 original sequence version: 106 4A0D0D56B5409D0A ikkb_human Inhibitor of nuclear factor kappa-B kinase subunit beta IKBKB IKKB I-kappa-B kinase 2 Nuclear factor NF-kappa-B inhibitor kinase beta Serine/threonine protein kinase IKBKB protein protein human Homo sapiens Human MSWSPSLTTQTCGAWEMKERLGTGGFGNVIRWHNQETGEQIAIKQCRQELSPRNRERWCLEIQIMRRLTHPNVVAARDVPEGMQNLAPNDLPLLAMEYCQGGDLRKYLNQFENCCGLREGAILTLLSDIASALRYLHENRIIHRDLKPENIVLQQGEQRLIHKIIDLGYAKELDQGSLCTSFVGTLQYLAPELLEQQKYTVTVDYWSFGTLAFECITGFRPFLPNWQPVQWHSKVRQKSEVDIVVSEDLNGTVKFSSSLPYPNNLNSVLAERLEKWLQLMLMWHPRQRGTDPTYGPNGCFKALDDILNLKLVHILNMVTGTIHTYPVTEDESLQSLKARIQQDTGIPEEDQELLQEAGLALIPDKPATQCISDGKLNEGHTLDMDLVFLFDNSKITYETQISPRPQPESVSCILQEPKRNLAFFQLRKVWGQVWHSIQTLKEDCNRLQQGQRAAMMNLLRNNSCLSKMKNSMASMSQQLKAKLDFFKTSIQIDLEKYSEQTEFGITSDKLLLAWREMEQAVELCGRENEVKLLVERMMALQTDIVDLQRSPMGRKQGGTLDDLEEQARELYRRLREKPRDQRTEGDSQEMVRLLLQAIQSFEKKVRVIYTQLSKTVVCKQKALELLPKVEEVVSLMNEDEKTVVRLQEKRQKELWNLLKIACSKVRGPVSGSPDSMNASRLSQPGQLMSQPSTASNSLPEPAKKSEELVAEAHNLCTLLENAIQDTVREQDQSFTALDWSWLQTEEEEHSCLEQAS DIP protein O14920 original sequence version: 89 DIP protein O14920 original sequence version: 100 DIP protein O14920 original sequence version: 121 DIP protein O14920 original sequence version: 126 DIP protein O14920 original sequence version: 136 DIP protein O14920 original sequence version: 128 DIP protein O14920-1 original sequence version: 151 DIP protein O14920 original sequence version: 142 DIP protein O14920 original sequence version: 159 DIP protein O14920 original sequence version: 93 DIP protein O14920 original sequence version: 124 DIP protein O14920-1 original sequence version: 178 DIP protein O14920 original sequence version: 85 F9CADF671AE9E14E ikkb_mouse Inhibitor of nuclear factor kappa-B kinase subunit beta I-kappa-B kinase 2 Nuclear factor NF-kappa-B inhibitor kinase beta Ikbkb Ikkb Serine/threonine protein kinase IKBKB protein protein mouse Mus musculus Mouse MSWSPSLPTQTCGAWEMKERLGTGGFGNVIRWHNQATGEQIAIKQCRQELSPKNRNRWCLEIQIMRRLNHPNVVAARDVPEGMQNLAPNDLPLLAMEYCQGGDLRRYLNQFENCCGLREGAVLTLLSDIASALRYLHENRIIHRDLKPENIVLQQGEKRLIHKIIDLGYAKELDQGSLCTSFVGTLQYLAPELLEQQKYTVTVDYWSFGTLAFECITGFRPFLPNWQPVQWHSKVRQKSEVDIVVSEDLNGAVKFSSSLPFPNNLNSVLAERLEKWLQLMLMWHPRQRGTDPQYGPNGCFRALDDILNLKLVHVLNMVTGTVHTYPVTEDESLQSLKTRIQENTGILETDQELLQKAGLVLLPDKPATQCISDSKTNEGLTLDMDLVFLLDNSKINYETQITPRPPPESVSCILQEPKRNLSFFQLRKVWGQVWHSIQTLKEDCNRLQQGQRAAMMSLLRNNSCLSKMKNAMASTAQQLKAKLDFFKTSIQIDLEKYKEQTEFGITSDKLLLAWREMEQAVEQCGRENDVKHLVERMMALQTDIVDLQRSPMGRKQGGTLDDLEEQARELYRKLREKPRDQRTEGDSQEMVRLLLQAIQSFEKKVRVIYTQLSKTVVCKQKALELLPKVEEVVSLMNEDERTVVRLQEKRQKELWNLLKIACSKVRGPVSGSPDSMNVSRLSHPGQLMSQPSSACDSLPESDKKSEELVAEAHALCSRLESALQDTVKEQDRSFTTLDWSWLQMEDEERCSLEQACD DIP protein O88351 original sequence version: 126 DIP protein O88351 original sequence version: 73 DIP protein O88351 original sequence version: 99 DIP protein O88351 original sequence version: 114 DIP protein O88351 original sequence version: 149 FED962F095449C5E snp23_mouse Synaptosomal-associated protein 23 Snap23 Sndt Vesicle-membrane fusion protein SNAP-23 Syndet protein protein mouse Mus musculus Mouse MDNLSPEEVQLRAHQVTDESLESTRRILGLAIESQDAGIKTITMLDEQGEQLNRIEEGMDQINKDMREAEKTLTELNKCCGLCICPCNRTKNFESGKNYKATWGDGGDNSPSNVVSKQPSRITNGQPQQTTGAASGGYIKRITNDAREDEMEENLTQVGSILGNLKNMALDMGNEIDAQNQQIQKITEKADTNKNRIDIANTRAKKLIDS DIP protein O09044 original sequence version: 110 6919E127E16BA2C9 snp23_rat Synaptosomal-associated protein 23 Snap23 Vesicle-membrane fusion protein SNAP-23 protein protein rat Rattus norvegicus (Rat) Rat MDDLSPEEIQLRAHQVTDESLESTRRILGLAIESQDAGIKTITMLDEQGEQLNRIEEGMDQINKDMREAEKTLTELNKCCGLCVCPCNRTKNFESGKNYKATWGDGGDSSPSNVVSKQPSRITNGQPQQTTGAASGGYIKRITNDAREDEMEENLTQVGSILGNLKNMALDMGNEIDAQNQQIQKITEKADTNKNRIDIANTRAKKLIDS 0D63E3A6F9FE3BA2 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 snap23-stx4-1 2 10 unspecified role unspecified role neutral component neutral component 14 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component physical association physical association Lipid rafts identified by the cross-linking of glycosphingolipid GM1 with choleratoxin B subunit (CTx) Fig. 4A SNAP-23 and syntaxin 4 were detected in the membrane raft fractions before and after Fcvar epsilonRI stimulation No snap23-stx4-2 2 8 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component 14 unspecified role unspecified role neutral component neutral component 10 unspecified role unspecified role neutral component neutral component physical association physical association Lipid rafts identified by the cross-linking of glycosphingolipid GM1 with choleratoxin B subunit (CTx) Fig. 4A SNAP-23 and syntaxin 4 were detected in the membrane raft fractions before and after Fcvar epsilonRI stimulation Yes snap23-ikbk-1 1 9 unspecified role unspecified role prey prey 14 unspecified role unspecified role bait bait physical association physical association Lipid rafts identified by the cross-linking of glycosphingolipid GM1 with choleratoxin B subunit (CTx) Fig. 4C IKK2 and SNAP-23 coimmunoprecipitated in the raft fractions following Fcvar epsilonRI stimulation (Figure 4C). FC epsilon RI stimulation ikbkb-snap23-1 4 13 enzyme target enzyme target substrate substrate 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 O09044:p.[Ser95Ala;Ser120Ala] Alters kinetics of interaction mutation mutation certain certain sequence position certain certain certain sequence position certain S A certain certain sequence position certain certain certain sequence position certain S A Wed May 02 17:43:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser95ala,ser120ala' Tue Jun 30 17:16:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser95Ala;Ser120Ala]' ecobb Escherichia coli (strain B / BL21) 11 enzyme enzyme 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 spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm phosphorylation phosphorylation reaction phosphorylation IKK2 Phosphorylates SNAP-23 on Ser120 and Ser95 Phosphorylation of SNAP-23 (O09044) on Ser120 and Ser95 Fig. 5A snap23-stx4-3 3 15 unspecified role unspecified role neutral component neutral component 13 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component 6 unspecified role unspecified role neutral component neutral component association association It thus appears that IKK2 regulates the formation of SNARE complex of SNAP-23/syntaxin 4/VAMP-2, resulting in the membrane fusions in the mast cells. Fig. 5D Interaction does not occur in IKBKB -/- mast cells Fcepsilon RI stimulation ikbkb-snap23-2 5 12 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 13 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association IKK2 associates with GFP-SNAP-23 when coexpressed in 293T cells by immunoprecipitation (IP) assay. Fig. 4B