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/ colotti-2006-3 in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting western blot western blot Immuno blot Immuno blot Colotti G., Zamparelli C., Verzili D., Mella M., Loughrey CM., Smith GL., Chiancone E. Cardiac - Interactions involving cardiac related proteins A Calcium binding Study emilia.chiancone@uniroma1.it. Biochemistry 2006 Only protein-protein interactions imex curation imex curation colotti-2006-2 in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Colotti G., Zamparelli C., Verzili D., Mella M., Loughrey CM., Smith GL., Chiancone E. Cardiac - Interactions involving cardiac related proteins A Calcium binding Study emilia.chiancone@uniroma1.it. Biochemistry 2006 Only protein-protein interactions imex curation imex curation colotti-2006-1 in vitro In vitro fluorescence spectr classical fluorescence spectroscopy fluorescence spectr predetermined predetermined participant predetermined Colotti G., Zamparelli C., Verzili D., Mella M., Loughrey CM., Smith GL., Chiancone E. Cardiac - Interactions involving cardiac related proteins Determination of Calcium affinity using a calcium chelator. emilia.chiancone@uniroma1.it. Biochemistry 2006 Only protein-protein interactions imex curation imex curation colotti-2006-4 in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Colotti G., Zamparelli C., Verzili D., Mella M., Loughrey CM., Smith GL., Chiancone E. Cardiac - Interactions involving cardiac related proteins emilia.chiancone@uniroma1.it. Biochemistry 2006 Only protein-protein interactions imex curation imex curation anxa7_mouse Annexin A7 Anxa7 Anx7 Annexin VII Annexin-7 Synexin protein protein mouse Mus musculus Mouse MSYPGYPPTGYPPFPGYPPAGQESSFPTAGQYPYPSGFPPMGGGAYPPAPSGGYPGAGGYPAPGGYPAPGGYPGALSPGGPPAYPGGQGFGAPPGGAGFSGYPQPPAQSYGGGPAQVPVPGGFPGGQMPSQYPGGQAPYPSQPASMTQGTQGTILPASNFDAMRDAEILRKAMKGFGTDEQAIVDVVSNRSNDQRQQIKAAFKTMYGKDLIKDLKSELSGNMEELILALFMPSTYYDAWSLRKAMQGAGTQERVLIEILCTRTNQEIRDIVRCYQLEFGRDLEKDIRSDTSGHFERLLVSMCQGNRDERQSVNHQMAQEDAQRLYQAGEGRLGTDESCFNMILATRSFPQLKATMEAYSRMANRDLLSSVSREFSGYVESGLKTILQCALNRPAFFAERLYYSMKGAGTDDSTLVRIVVTRSEIDLVQIKQMFTQMYQKTLSTMIASDTSGDYRKLLLAIVGQ sorcn_crigr Sorcin 22 kDa protein CP-22 V19 SRI protein protein crigr Cricetulus griseus (Chinese hamster) Cricetulus barabensis griseus Chinese hamster MAYPGHPGAGGGYYPGGYGGAPGGPSFPGQTQDPLYGYFASVAGQDGQIDADELQRCLTQSGIAGGYKPFNLETCRLMVSMLDRDMSGTMGFNEFKELWAVLNGWRQHFISFDSDRSGTVDPQELQKALTTMGFRLNPQTVNSIAKRYSTSGKITFDDYIACCVKLRALTDSFRRRDSAQQGMVNFSYDDFIQCVMTV calcium ion calcium ion calcium ion calcium ions small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) g3i2p8_crigr Annexin I79_017691 protein protein crigr Cricetulus griseus (Chinese hamster) Cricetulus barabensis griseus Chinese hamster MSYPGYPPTGYPPFPGYPPAGQESSFPPPGQYPYPSGFPPMGGGAYPPAPSVGYPGAGGYATPGGYPAPGGYPVSPSPGGVPVYPGGTDEQAIVDVVSSRSNDQRQQIKTAFKTMYGKDLIKDLKSELSGNMEELILALFMPSTYYDAWSLRKAMQGAGTQERVLIEILCTRTNQEIRDIVRCYQSEFGRDLEKDIKSDTSGHFERLLVSMCQIDLVQIKQMFTQMYQKTLSTMIASDTSGDYRKLLLAIVGQ anxa7-sri-1 1796419 N/A 1796423 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) N/A 1796424 unspecified role unspecified role prey prey P05044:p.Trp99Gly 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 Figure 4A Interaction is calcium-dependent. anxa7-sri-2 1796419 N/A 1796423 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) N/A 1796424 unspecified role unspecified role prey prey P05044:p.Trp105Gly 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 Figure 4B Interaction is calcium-dependent. sri-9 1796420 N/A 1796425 unspecified role unspecified role neutral component neutral component N/A 1796424 unspecified role unspecified role neutral component neutral component P05044:p.Trp99Gly 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 Figure 3A When the level of calcium exceeds 2 equiv/monomer Sorcin formed aggregates. calcium ion-sri-2 1796421 N/A 1796425 unspecified role unspecified role neutral component neutral component N/A 1796424 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figure 3B Ca2+ affinity of the mutants is practically unaltered relative to that of wt sorcin and can be fitted with two Ca2+ binding constants in the micromolar range [K1 = (0.5 ± 0.2) × 10-6 M and K2 ∼ 10-5 M]. anxa7-sri-3 1796422 N/A 1796426 unspecified role unspecified role bait bait N/A sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 1796424 unspecified role unspecified role prey prey P05044:p.Trp105Gly mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 1796422 Figure 5 The W99G variant behaves like wt sorcin at <40 uM Ca2+ but yields lower Req values at higher cation concentrations. Kd values are entered for results obtained using 20uM calcium.