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/ The W105G and W99G sorcin mutants demonstrate the role of the D helix in the Ca(2+)-dependent interaction with annexin VII and the cardiac ryanodine receptor. The W105G and W99G sorcin mutants demonstrate the role of the D helix in the Ca(2+)-dependent interaction with annexin VII and the cardiac ryanodine receptor. Biochemistry 2006 imex curation Colotti G., Zamparelli C., Verzili D., Mella M., Loughrey CM., Smith GL., Chiancone E. Cardiac - Interactions involving cardiac related proteins emilia.chiancone@uniroma1.it. Only protein-protein interactions imex curation 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. Accepted 2015-JUN-02 by ORCHARD Biochemistry 2006 Only protein-protein interactions imex curation imex curation The Annexin 7 used is described above Figure 4 as given in Refs 15 (the pmid you give) and also Ref 31 (pmid: 9268363). However Ref 31 describes a Human protein - N terminal region 1-145 + GST tag. I have altered to Mouse, as you suggest, but not sure if this is what was actually used.; Comments removed and species of Anaz7 altered. As I pointed out the Kd for the W105G mutant is already present, and the difference between WT and W99G mutant is 0.1 so I did not enter this mutant at all. The W105G and W99G sorcin mutants demonstrate the role of the D helix in the Ca(2+)-dependent interaction with annexin VII and the cardiac ryanodine receptor. The W105G and W99G sorcin mutants demonstrate the role of the D helix in the Ca(2+)-dependent interaction with annexin VII and the cardiac ryanodine receptor. Biochemistry 2006 imex curation Colotti G., Zamparelli C., Verzili D., Mella M., Loughrey CM., Smith GL., Chiancone E. Cardiac - Interactions involving cardiac related proteins emilia.chiancone@uniroma1.it. Only protein-protein interactions imex curation 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. Accepted 2015-JUN-02 by ORCHARD Biochemistry 2006 Only protein-protein interactions imex curation imex curation The Annexin 7 used is described above Figure 4 as given in Refs 15 (the pmid you give) and also Ref 31 (pmid: 9268363). However Ref 31 describes a Human protein - N terminal region 1-145 + GST tag. I have altered to Mouse, as you suggest, but not sure if this is what was actually used.; Comments removed and species of Anaz7 altered. As I pointed out the Kd for the W105G mutant is already present, and the difference between WT and W99G mutant is 0.1 so I did not enter this mutant at all. The W105G and W99G sorcin mutants demonstrate the role of the D helix in the Ca(2+)-dependent interaction with annexin VII and the cardiac ryanodine receptor. The W105G and W99G sorcin mutants demonstrate the role of the D helix in the Ca(2+)-dependent interaction with annexin VII and the cardiac ryanodine receptor. Biochemistry 2006 imex curation Colotti G., Zamparelli C., Verzili D., Mella M., Loughrey CM., Smith GL., Chiancone E. Cardiac - Interactions involving cardiac related proteins emilia.chiancone@uniroma1.it. Only protein-protein interactions imex curation 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 I am not sure what this expt is meant to be showing - A Calcium binding Study emilia.chiancone@uniroma1.it. Accepted 2015-JUN-02 by ORCHARD Biochemistry 2006 Only protein-protein interactions imex curation imex curation The Annexin 7 used is described above Figure 4 as given in Refs 15 (the pmid you give) and also Ref 31 (pmid: 9268363). However Ref 31 describes a Human protein - N terminal region 1-145 + GST tag. I have altered to Mouse, as you suggest, but not sure if this is what was actually used.; Comments removed and species of Anaz7 altered. As I pointed out the Kd for the W105G mutant is already present, and the difference between WT and W99G mutant is 0.1 so I did not enter this mutant at all. The W105G and W99G sorcin mutants demonstrate the role of the D helix in the Ca(2+)-dependent interaction with annexin VII and the cardiac ryanodine receptor. The W105G and W99G sorcin mutants demonstrate the role of the D helix in the Ca(2+)-dependent interaction with annexin VII and the cardiac ryanodine receptor. Biochemistry 2006 imex curation Colotti G., Zamparelli C., Verzili D., Mella M., Loughrey CM., Smith GL., Chiancone E. Cardiac - Interactions involving cardiac related proteins emilia.chiancone@uniroma1.it. Only protein-protein interactions imex curation 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. Accepted 2015-JUN-02 by ORCHARD Biochemistry 2006 Only protein-protein interactions imex curation imex curation The Annexin 7 used is described above Figure 4 as given in Refs 15 (the pmid you give) and also Ref 31 (pmid: 9268363). However Ref 31 describes a Human protein - N terminal region 1-145 + GST tag. I have altered to Mouse, as you suggest, but not sure if this is what was actually used.; Comments removed and species of Anaz7 altered. As I pointed out the Kd for the W105G mutant is already present, and the difference between WT and W99G mutant is 0.1 so I did not enter this mutant at all. g3i2p8_crigr Annexin I79_017691 protein protein crigr Cricetulus griseus (Chinese hamster) Cricetulus barabensis griseus Chinese hamster MSYPGYPPTGYPPFPGYPPAGQESSFPPPGQYPYPSGFPPMGGGAYPPAPSVGYPGAGGYATPGGYPAPGGYPVSPSPGGVPVYPGGTDEQAIVDVVSSRSNDQRQQIKTAFKTMYGKDLIKDLKSELSGNMEELILALFMPSTYYDAWSLRKAMQGAGTQERVLIEILCTRTNQEIRDIVRCYQSEFGRDLEKDIKSDTSGHFERLLVSMCQIDLVQIKQMFTQMYQKTLSTMIASDTSGDYRKLLLAIVGQ 4E0491154228C04B anxa7_mouse Annexin A7 Anxa7 Anx7 Annexin VII Annexin-7 Synexin protein protein mouse Mus musculus Mouse MSYPGYPPTGYPPFPGYPPAGQESSFPTAGQYPYPSGFPPMGGGAYPPAPSGGYPGAGGYPAPGGYPAPGGYPGALSPGGPPAYPGGQGFGAPPGGAGFSGYPQPPAQSYGGGPAQVPVPGGFPGGQMPSQYPGGQAPYPSQPASMTQGTQGTILPASNFDAMRDAEILRKAMKGFGTDEQAIVDVVSNRSNDQRQQIKAAFKTMYGKDLIKDLKSELSGNMEELILALFMPSTYYDAWSLRKAMQGAGTQERVLIEILCTRTNQEIRDIVRCYQLEFGRDLEKDIRSDTSGHFERLLVSMCQGNRDERQSVNHQMAQEDAQRLYQAGEGRLGTDESCFNMILATRSFPQLKATMEAYSRMANRDLLSSVSREFSGYVESGLKTILQCALNRPAFFAERLYYSMKGAGTDDSTLVRIVVTRSEIDLVQIKQMFTQMYQKTLSTMIASDTSGDYRKLLLAIVGQ 73339650748B4689 sorcn_crigr Sorcin 22 kDa protein CP-22 V19 SRI protein protein crigr Cricetulus griseus (Chinese hamster) Cricetulus barabensis griseus Chinese hamster MAYPGHPGAGGGYYPGGYGGAPGGPSFPGQTQDPLYGYFASVAGQDGQIDADELQRCLTQSGIAGGYKPFNLETCRLMVSMLDRDMSGTMGFNEFKELWAVLNGWRQHFISFDSDRSGTVDPQELQKALTTMGFRLNPQTVNSIAKRYSTSGKITFDDYIACCVKLRALTDSFRRRDSAQQGMVNFSYDDFIQCVMTV DA2AF8A9707561C5 calcium ion calcium ion calcium ion calcium ions small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) sri-9 3 8 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component P05044:p.Trp99Gly mutation mutation certain certain sequence position certain certain certain sequence position certain W G Wed May 02 18:38:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp99gly' Tue Jun 30 17:59:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp99Gly' ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 3A When the level of calcium exceeds 2 equiv/monomer Sorcin formed aggregates. calcium ion-sri-2 2 8 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction 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-1 1 6 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey P05044:p.Trp99Gly mutation mutation certain certain sequence position certain certain certain sequence position certain W G Wed May 02 16:29:23 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp99gly' Tue Jun 30 20:12:26 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp99Gly' ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 4A Interaction is calcium-dependent. anxa7-sri-2 1 6 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey P05044:p.Trp105Gly mutation mutation certain certain sequence position certain certain certain sequence position certain W G Wed May 02 16:13:03 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp105gly' Tue Jun 30 16:43:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp105Gly' ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 4B Interaction is calcium-dependent. anxa7-sri-3 4 5 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 chemical synthesis Chemical synthesis (Chemical synthesis) 7 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 W G Wed May 02 19:58:10 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp105gly' Tue Jun 30 16:55:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp105Gly' ecoli Escherichia coli (strain K12) direct interaction direct interaction 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. I have added one Kd for both WT and W99G variant and a separate one for the 105 variant.