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/ lovell-2008-1 Membrane Binding by tBid Initiates an Ordered Series of Events Culminating in Membrane Permeabilization by Bax. in vitro In vitro fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Lovell JF., Billen LP., Bindner S., Shamas-Din A., Fradin C., Leber B., Andrews DW. Apoptosis - Interactions involving proteins with a function related to apoptosis Assays were performed in the presence of liposomes, into which the proteins are assumed to have inserted. Liposomes wee then lysed and proteins separated on a Superdex 200 column. Source of BAX given in PMID:15920484 referenced by the author andrewsd@mcmaster.ca Cell (0092-8674) 2008 Only protein-protein interactions 4 imex curation lovell-2008-2 Membrane Binding by tBid Initiates an Ordered Series of Events Culminating in Membrane Permeabilization by Bax. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Lovell JF., Billen LP., Bindner S., Shamas-Din A., Fradin C., Leber B., Andrews DW. Apoptosis - Interactions involving proteins with a function related to apoptosis Assays were performed in the presence of liposomes, into which the proteins are assumed to have inserted. Liposomes wee then lysed and proteins separated on a Superdex 200 column. Source of BAX given in PMID:15920484 referenced by the author andrewsd@mcmaster.ca Cell (0092-8674) 2008 Only protein-protein interactions 4 imex curation p70444-pro_0000223236 BH3-interacting domain death agonist p15 Bid p22 BID protein protein mouse Mus musculus Mouse SQASRSFNQGRIEPDSESQEEIIHNIARHLAQIGDEMDHNIQPTLVRQLAAQFMNGSLSEEDKRNCLAKALDEVKTAFPRDMENDKAMLIMTMLLAKKVASHAPSLLRDVFHTTVNFINQNLFSYVRNLVRNEMD 61 195 bax_human Apoptosis regulator BAX BAX BCL2L4 Bcl-2-like protein 4 protein protein human Homo sapiens Human MDGSGEQPRGGGPTSSEQIMKTGALLLQGFIQDRAGRMGGEAPELALDPVPQDASTKKLSECLKRIGDELDSNMELQRMIAAVDTDSPREVFFRVAADMFSDGNFNWGRVVALFYFASKLVLKALCTKVPELIRTIMGWTLDFLRERLLGWIQDQGGWDGLLSYFGTPTWQTVTIFVAGVLTASLTIWKKMG bid_mouse BH3-interacting domain death agonist p22 BID Bid protein protein mouse Mus musculus Mouse MDSEVSNGSGLGAEHITDLLVFGFLQSSGCTRQELEVLGRELPVQAYWEADLEDELQTDGSQASRSFNQGRIEPDSESQEEIIHNIARHLAQIGDEMDHNIQPTLVRQLAAQFMNGSLSEEDKRNCLAKALDEVKTAFPRDMENDKAMLIMTMLLAKKVASHAPSLLRDVFHTTVNFINQNLFSYVRNLVRNEMD o-phosphoethanolamine Phosphoethanolamine small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) bax-tbid-1 Interaction of BAX and tBid demonstrated by FRET 783399 n/a 783401 unspecified role unspecified role fluorescence donor fluorescence donor P70444-PRO_0000223236:p.Gly34Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false cys-126 dimethylaminocoumarin (DAC) fluorescent dye fluorescent dye label fluorescent dye certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 783402 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept cys-126 7-nitrobenzene-2-oxa-1, 3-diazol-4-yl ethylenediamine fluorescent dye fluorescent dye label fluorescent dye certain certain sequence position certain certain certain sequence position certain false Q07812:p.Cys62Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figs. 2A, 2C, 4 Energy transfer was observed only if liposomes were also present, suggesting that the two proteins interact only when bound to membranes. pH = 7.0, Temp = 37C bax-tbid-2 Interaction of BAX and tBid demonstrated by FRET, inhibition by Bid BH3 peptide 783399 n/a 783402 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept cys-126 7-nitrobenzene-2-oxa-1, 3-diazol-4-yl ethylenediamine fluorescent dye fluorescent dye label fluorescent dye certain certain sequence position certain certain certain sequence position certain false Q07812:p.Cys62Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 783403 inhibitor inhibitor ancillary ancillary bh3 region sufficient to bind sufficient binding region sufficient to bind range range range range false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 783401 unspecified role unspecified role fluorescence donor fluorescence donor cys-126 dimethylaminocoumarin (DAC) fluorescent dye fluorescent dye label fluorescent dye certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false Fig. 2B Exact coordinates of Bid peptide not given in paper so have been inferred from the UniProtKB entry. Energy transfer was observed only if liposomes were also present, suggesting that the two proteins interact only when bound to membranes. tbid-pe-1 Binding of tBid to phosphoethanolamine demonstrated by FRET 783399 n/a 783404 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept n/a 783401 unspecified role unspecified role fluorescence donor fluorescence donor P70444-PRO_0000223236:p.Gly34Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false cys-126 dimethylaminocoumarin (DAC) fluorescent dye fluorescent dye label fluorescent dye certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false Fig. 3A Phosphoethanolomaine is tagged with a fluorescence dye to form 1-Oleoyl-2-[12-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]dodecanoyl]-sn-Glycero-3-Phosphoethanolamine (NBD-PE) bax-1 Homo-oligermerisation of BAX 783399 n/a 783402 unspecified role unspecified role fluorescence donor fluorescence donor Q07812:p.Arg134Cys mutation mutation certain certain sequence position certain certain certain sequence position certain false cys-134 dimethylaminocoumarin (DAC) fluorescent dye fluorescent dye label fluorescent dye certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 783402 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept cys-126 7-nitrobenzene-2-oxa-1, 3-diazol-4-yl ethylenediamine fluorescent dye fluorescent dye label fluorescent dye certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Energy transfer was observed only if liposomes were also present, suggesting that the two proteins interact only when bound to membranes. Fig. 3B bax-5 Homo-dimerisation of BAX 783400 n/a 783402 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Interaction requires the presence of a liosome membrane preparation. Supp. Fig. 1B