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/ soundar-2003-1 Evaluation by mutagenesis of the importance of 3 arginines in alpha, beta, and gamma subunits of human NAD-dependent isocitrate dehydrogenase. 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 J. Biol. Chem. (0021-9258) 2003 Only protein-protein interactions imex curation soundar-2003-2 Evaluation by mutagenesis of the importance of 3 arginines in alpha, beta, and gamma subunits of human NAD-dependent isocitrate dehydrogenase. 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 J. Biol. Chem. (0021-9258) 2003 Only protein-protein interactions imex curation soundar-2003-3 Evaluation by mutagenesis of the importance of 3 arginines in alpha, beta, and gamma subunits of human NAD-dependent isocitrate dehydrogenase. 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 J. Biol. Chem. (0021-9258) 2003 Only protein-protein interactions imex curation soundar-2003-4 Evaluation by mutagenesis of the importance of 3 arginines in alpha, beta, and gamma subunits of human NAD-dependent isocitrate dehydrogenase. 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 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 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 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 adp adenosine 5'-(trihydrogen diphosphate) (ADP) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) idh3a-idh3b-1 Components of the Isocitrate Dehydrogenase Holoenzyme identified by Edman degradation 596408 N/A 596412 unspecified role unspecified role neutral component neutral component N/A 596413 unspecified role unspecified role neutral component neutral component N/A 596414 unspecified role unspecified role neutral component neutral component physical association physical association false false false 2 idh3a-idh3b-2 Components of the Isocitrate Dehydrogenase Holoenzyme identified by gel filtration 596409 N/A 596412 unspecified role unspecified role neutral component neutral component N/A 596414 unspecified role unspecified role neutral component neutral component N/A 596413 unspecified role unspecified role neutral component neutral component physical association physical association false false false 3 Molecular mass of the complex is approximately 315kDa, indicating an octomer. idh3a-idh3b-3 Components of the Isocitrate Dehydrogenase Holoenzyme identified by gel filtration 596410 N/A 596414 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 N/A 596413 unspecified role unspecified role neutral component neutral component Stoichiometry: 2.0 N/A 596412 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 physical association physical association false false false 4 idh3a-idh3b-4 Components of the Isocitrate Dehydrogenase Holoenzyme identified by ultrafiltration 596411 N/A 596413 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 false Stoichiometry: 2.0 N/A 596412 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 false Stoichiometry: 1.0 N/A 596415 unspecified role unspecified role neutral component neutral component N/A 596414 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 false Stoichiometry: 1.0 physical association physical association false false false 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.