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/ Evaluation by mutagenesis of the importance of 3 arginines in alpha, beta, and gamma subunits of human NAD-dependent isocitrate dehydrogenase. Evaluation by mutagenesis of the importance of 3 arginines in alpha, beta, and gamma subunits of human NAD-dependent isocitrate dehydrogenase. J. Biol. Chem. (0021-9258) 2003 imex curation Soundar S., Park JH., Huh TL., Colman RF. Enzymes expressed in E.coli from a single vector. Complex purified by ammonium sulfate fractionation and ion exchange chromatography. Complex composition was examined by SDS-PAGE followed by Edman sequencing of the three subunits. rfcolman@udel.edu 10-APR-2007: Contacted by JYOTI. Only protein-protein interactions ecoli Escherichia coli (strain K12) ion exchange chrom ion exchange chromatography IEC IEC ion exchange chrom edman degradation edman degradation Soundar S., Park JH., Huh TL., Colman RF. Enzymes expressed in E.coli from a single vector. Complex purified by ammonium sulfate fractionation and ion exchange chromatography. Complex composition was examined by SDS-PAGE followed by Edman sequencing of the three subunits. rfcolman@udel.edu Accepted 2007-APR-23 by JYOTI. 10-APR-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2003 Only protein-protein interactions imex curation Evaluation by mutagenesis of the importance of 3 arginines in alpha, beta, and gamma subunits of human NAD-dependent isocitrate dehydrogenase. Evaluation by mutagenesis of the importance of 3 arginines in alpha, beta, and gamma subunits of human NAD-dependent isocitrate dehydrogenase. J. Biol. Chem. (0021-9258) 2003 imex curation Soundar S., Park JH., Huh TL., Colman RF. Enzymes expressed in E.coli from a single vector. Complex purified by ammonium sulfate fractionation and ion exchange chromatography. Complex composition was examined by SDS-PAGE followed by Edman sequencing of the three subunits. rfcolman@udel.edu 10-APR-2007: Contacted by JYOTI. Only protein-protein interactions ecoli Escherichia coli (strain K12) solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Soundar S., Park JH., Huh TL., Colman RF. Enzymes expressed in E.coli from a single vector. Complex purified by ammonium sulfate fractionation and ion exchange chromatography. Complex composition was examined by SDS-PAGE followed by Edman sequencing of the three subunits. rfcolman@udel.edu Accepted 2007-APR-23 by JYOTI. 10-APR-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2003 Only protein-protein interactions imex curation Evaluation by mutagenesis of the importance of 3 arginines in alpha, beta, and gamma subunits of human NAD-dependent isocitrate dehydrogenase. Evaluation by mutagenesis of the importance of 3 arginines in alpha, beta, and gamma subunits of human NAD-dependent isocitrate dehydrogenase. J. Biol. Chem. (0021-9258) 2003 imex curation Soundar S., Park JH., Huh TL., Colman RF. Enzymes expressed in E.coli from a single vector. Complex purified by ammonium sulfate fractionation and ion exchange chromatography. Complex composition was examined by SDS-PAGE followed by Edman sequencing of the three subunits. rfcolman@udel.edu 10-APR-2007: Contacted by JYOTI. Only protein-protein interactions ecoli Escherichia coli (strain K12) reverse phase chrom reverse phase chromatography reverse phase chrom edman degradation edman degradation Soundar S., Park JH., Huh TL., Colman RF. Enzymes expressed in E.coli from a single vector. Complex purified by ammonium sulfate fractionation and ion exchange chromatography. Complex composition was examined by SDS-PAGE followed by Edman sequencing of the three subunits. rfcolman@udel.edu Accepted 2007-APR-23 by JYOTI. 10-APR-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2003 Only protein-protein interactions imex curation Evaluation by mutagenesis of the importance of 3 arginines in alpha, beta, and gamma subunits of human NAD-dependent isocitrate dehydrogenase. Evaluation by mutagenesis of the importance of 3 arginines in alpha, beta, and gamma subunits of human NAD-dependent isocitrate dehydrogenase. J. Biol. Chem. (0021-9258) 2003 imex curation Soundar S., Park JH., Huh TL., Colman RF. Enzymes expressed in E.coli from a single vector. Complex purified by ammonium sulfate fractionation and ion exchange chromatography. Complex composition was examined by SDS-PAGE followed by Edman sequencing of the three subunits. rfcolman@udel.edu 10-APR-2007: Contacted by JYOTI. Only protein-protein interactions ecoli Escherichia coli (strain K12) molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Soundar S., Park JH., Huh TL., Colman RF. Enzymes expressed in E.coli from a single vector. Complex purified by ammonium sulfate fractionation and ion exchange chromatography. Complex composition was examined by SDS-PAGE followed by Edman sequencing of the three subunits. rfcolman@udel.edu Accepted 2007-APR-23 by JYOTI. 10-APR-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2003 Only protein-protein interactions imex curation idh3g_human Isocitrate dehydrogenase [NAD] subunit gamma, mitochondrial IDH3G Isocitric dehydrogenase subunit gamma NAD(+)-specific ICDH subunit gamma protein protein human Homo sapiens Human MALKVATVAGSAAKAVLGPALLCRPWEVLGAHEVPSRNIFSEQTIPPSAKYGGRHTVTMIPGDGIGPELMLHVKSVFRHACVPVDFEEVHVSSNADEEDIRNAIMAIRRNRVALKGNIETNHNLPPSHKSRNNILRTSLDLYANVIHCKSLPGVVTRHKDIDILIVRENTEGEYSSLEHESVAGVVESLKIITKAKSLRIAEYAFKLAQESGRKKVTAVHKANIMKLGDGLFLQCCREVAARYPQITFENMIVDNTTMQLVSRPQQFDVMVMPNLYGNIVNNVCAGLVGGPGLVAGANYGHVYAVFETATRNTGKSIANKNIANPTATLLASCMMLDHLKLHSYATSIRKAVLASMDNENMHTPDIGGQGTTSEAIQDVIRHIRVINGRAVEA A0870F2B7D228A37 adp adenosine 5'-(trihydrogen diphosphate) (ADP) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) idh3a_human Isocitrate dehydrogenase [NAD] subunit alpha, mitochondrial IDH3A Isocitric dehydrogenase subunit alpha NAD(+)-specific ICDH subunit alpha protein protein human Homo sapiens Human MAGPAWISKVSRLLGAFHNPKQVTRGFTGGVQTVTLIPGDGIGPEISAAVMKIFDAAKAPIQWEERNVTAIQGPGGKWMIPSEAKESMDKNKMGLKGPLKTPIAAGHPSMNLLLRKTFDLYANVRPCVSIEGYKTPYTDVNIVTIRENTEGEYSGIEHVIVDGVVQSIKLITEGASKRIAEFAFEYARNNHRSNVTAVHKANIMRMSDGLFLQKCREVAESCKDIKFNEMYLDTVCLNMVQDPSQFDVLVMPNLYGDILSDLCAGLIGGLGVTPSGNIGANGVAIFESVHGTAPDIAGKDMANPTALLLSAVMMLRHMGLFDHAARIEAACFATIKDGKSLTKDLGGNAKCSDFTEEICRRVKDLD 695F6A34F97430CF o43837-2 Isocitrate dehydrogenase [NAD] subunit beta, mitochondrial IDH3B Isocitric dehydrogenase subunit beta NAD(+)-specific ICDH subunit beta protein protein human Homo sapiens Human MAALSGVRWLTRALVSAGNPGAWRGLSTSAAAHAASRSQAEDVRVEGSFPVTMLPGDGVGPELMHAVKEVFKAAAVPVEFQEHHLSEVQNMASEEKLEQVLSSMKENKVAIIGKIHTPMEYKGELASYDMRLRRKLDLFANVVHVKSLPGYMTRHNNLDLVIIREQTEGEYSSLEHESARGVIECLKIVTRAKSQRIAKFAFDYATKKGRGKVTAVHKANIMKLGDGLFLQCCEEVAELYPKIKFETMIIDNCCMQLVQNPYQFDVLVMPNLYGNIIDNLAAGLVGGAGVVPGESYSAEYAVFETGARHPFAQAVGRNIANPTAMLLSASNMLRHLNLEYHSSMIADAVKKVIKVGKVRTSDMGGYATCHDFTEAVIAALPHP 6B5A3D3F82CCDB34 idh3a-idh3b-1 1 5 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component physical association physical association Edman degradation of each of the enzyme preparations reveals the presence of only the sequences of the {alpha}, {beta}, and {gamma} subunits of human NAD-dependent isocitrate dehydrogenase 2 idh3a-idh3b-2 4 5 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component physical association physical association Gel filtration of purified wild type and mutant enzymes. 3 Molecular mass of the complex is approximately 315kDa, indicating an octomer. idh3a-idh3b-3 3 8 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component 5 unspecified role unspecified role neutral component neutral component physical association physical association Determination of Subunit Composition of Enzymes by Reverse Phase Chromatography 4 idh3a-idh3b-4 2 7 unspecified role unspecified role neutral component neutral component P50213:p.Arg115Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R Q Wed May 02 16:20:09 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg115gln' Tue Jun 30 16:20:10 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg115Gln' 5 unspecified role unspecified role neutral component neutral component P51553:p.Arg136Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R Q Wed May 02 19:39:04 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg136gln' Tue Jun 30 15:37:51 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg136Gln' 6 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component O43837-2:p.Arg133Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R Q Wed May 02 20:09:03 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg133gln' Tue Jun 30 16:28:14 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg133Gln' physical association physical association ADP binding of wild type and mutant isocitrate dehydrogenases Table 4 The dissociation constant for enzyme-ADP is 15.9, 73.4, 234, and 513uM for wild type, alpha-R115, beta-R133Q, and gamma-R136Q enzymes, respectively.