1.1.1.87 additional information - additional information Michaelis-Menten kinetics, the enzyme shows a steady-state random kinetic mechanism with a preferred order of addition of Mg2+ prior to NAD+. The same step(s) limit the reaction at limiting and saturating Mg2+ concentrations. Solvent kinetic deuterium isotope effects and viscosity effects are consistent with a rate-limiting pre-catalytic conformational change at saturating reactant concentrations 739958 1.1.1.87 additional information - additional information mutant enzymes kinetic analysis and pH-dependencies, overview 696350 1.1.1.87 additional information - additional information substitution of potassium acetate for KCl changes the kinetic mechanism of HIcDH from a steady state random to a fully ordered mechanism with the binding of Mg-HIc followed by K+ and NAD+, increase in the affinity of enzyme for Mg-HIc as a result of elimination of the inhibitory effect of Cl-, kinetic analysis, overview 696220 1.1.1.87 0.0042 - homoisocitrate oxidate decarboxylation of homoisocitrate 685232 1.1.1.87 0.0073 - homoisocitrate wild type enzyme, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.01 - homoisocitrate value below 644539 1.1.1.87 0.014 - isocitrate wild type enzyme, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.0164 - isocitrate pH 7.8, 70°C, recombinant enzyme 660907 1.1.1.87 0.018 - homoisocitrate - 685539 1.1.1.87 0.018 - homoisocitrate pH 7.8, 36°C, recombinant enzyme 667934 1.1.1.87 0.0183 - homoisocitrate pH 7.8, 70°C, recombinant enzyme 660907 1.1.1.87 0.02 - trisodium (2S,3R)-2-(carboxylatomethoxy)-3-hydroxybutanedioate - 685539 1.1.1.87 0.036 - NAD+ wild type enzyme, with homoisocitrate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.037 - homoisocitrate mutant enzyme S80A, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.042 - homoisocitrate mutant enzyme L81P, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.042 - NAD+ native enzyme, pH 7.8, 20°C 722303 1.1.1.87 0.05 - 3-isopropylmalate mutant enzyme S80A, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.064 - homoisocitrate wild type enzyme, with NADP+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.065 - NADH - 644539 1.1.1.87 0.074 - (1R,2S)-1-hydroxybutane-1,2,4-tricarboxylate recombinant His-tagged enzyme, pH 7.8, 20°C 722303 1.1.1.87 0.083 - 3-vinylmalate pH 7.8, 28°C, recombinant enzyme 667934 1.1.1.87 0.09 - NADH reductive carboxylation of alpha-ketoadipate 685232 1.1.1.87 0.1 - NAD+ wild type enzyme, with isocitrate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.15 - trisodium (2S,3R)-2-[(carboxylatomethyl)amino]-3-hydroxybutanedioate - 685539 1.1.1.87 0.16 - NAD+ mutant enzyme L83R, with homoisocitrate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.161 - isocitrate pH 7.5, 60°C, recombinant mutant V135M 669109 1.1.1.87 0.17 - homoisocitrate mutant enzyme L83R, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.211 - homoisocitrate pH 7.8, 25°C, recombinant wild-type enzyme 660918 1.1.1.87 0.211 - homoisocitrate pH 7.8, 28°C, recombinant enzyme 667934 1.1.1.87 0.25 - NAD+ mutant enzyme T71V, with isocitrate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.29 - (2R,3S)-3-propylmalic acid pH 7.8, 36°C, recombinant enzyme 667934 1.1.1.87 0.291 - isocitrate pH 7.8, 25°C, recombinant wild-type enzyme 660918 1.1.1.87 0.291 - isocitrate pH 7.8, 28°C, recombinant enzyme 667934 1.1.1.87 0.3 - isocitrate mutant enzyme S80A, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.3 - NAD+ pH 8.0, 25°C, in presence of K+ 696220 1.1.1.87 0.31 - 3-isopropylmalate wild type enzyme, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.32 - NAD+ mutant enzyme L81P, with homoisocitrate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.33 - NAD+ - 644539 1.1.1.87 0.4 - 1-hydroxy-1,2,3-propanetricarboxylate purified enzyme, pH 8.0, 60°C 656127 1.1.1.87 0.41 - (2R,3S)-3-allylmalic acid pH 7.8, 36°C, recombinant enzyme 667934 1.1.1.87 0.44 - (2R,3S)-3-(4-hydroxybutyl)malate pH 7.8, 36°C, recombinant enzyme 667934 1.1.1.87 0.45 - (1R,2S)-1-hydroxybutane-1,2,4-tricarboxylate native enzyme, pH 7.8, 20°C 722303 1.1.1.87 0.45 - NAD+ oxidate decarboxylation of homoisocitrate 685232 1.1.1.87 0.46 - homoisocitrate mutant enzyme I82N, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.465 - isocitrate pH 7.8, 25°C, recombinant mutant A80del 660918 1.1.1.87 0.47 - (2R,3S)-3-(4-hydroxybutyl)malate pH 7.8, 28°C, recombinant enzyme 667934 1.1.1.87 0.5 - isocitrate mutant enzyme T71V, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.51 - (2R,3S)-3-(3-hydroxypropyl)malate pH 7.8, 28°C, recombinant enzyme 667934 1.1.1.87 0.521 - isocitrate pH 7.5, 60°C, recombinant wild-type enzyme 669109 1.1.1.87 0.58 - (2R,3S)-3-(3-hydroxypropyl)malate pH 7.8, 36°C, recombinant enzyme 667934 1.1.1.87 0.58 - isocitrate mutant enzyme L83R, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.59 - (2R,3S)-3-(2-hydroxyethyl)malate pH 7.8, 28°C, recombinant enzyme 667934 1.1.1.87 0.6 - ethylmalate pH 7.8, 36°C, recombinant enzyme 667934 1.1.1.87 0.6 - NAD+ mutant enzyme L81P, with isocitrate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.62 - NAD+ mutant enzyme L83R, with isocitrate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.64 - NAD+ mutant enzyme I82N, with homoisocitrate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.68 - 3-isopropylmalate mutant enzyme L81P, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.71 - 3-isopropylmalate mutant enzyme T71V, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.76 - NAD+ mutant enzyme S80A, with isocitrate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.78 - (2R,3S)-3-propylmalic acid pH 7.8, 28°C, recombinant enzyme 667934 1.1.1.87 0.819 - homoisocitrate pH 7.8, 25°C, recombinant mutant A80del 660918 1.1.1.87 0.92 - NAD+ mutant enzyme S80A, with homoisocitrate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.92 - NAD+ wild type enzyme, with D-malate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 0.94 - homoisocitrate mutant enzyme T71V, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 1 - ethylmalate pH 7.8, 28°C, recombinant enzyme 667934 1.1.1.87 1 - homoisocitrate pH 7.8, 25°C, recombinant mutant R87V 660918 1.1.1.87 1 - NAD+ mutant enzyme T71V, with 3-isopropylmalate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 1.09 - NAD+ recombinant His-tagged enzyme, pH 7.8, 20°C 722303 1.1.1.87 1.2 - isocitrate mutant enzyme L81P, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 1.2 - NAD+ mutant enzyme S80A, with 3-isopropylmalate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 1.3 - NAD+ mutant enzyme L81P, with 3-isopropylmalate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 1.33 - 3-isopropylmalate pH 7.8, 25°C, recombinant wild-type enzyme 660918 1.1.1.87 1.33 - isopropylmalate pH 7.8, 28°C, recombinant enzyme 667934 1.1.1.87 1.4 - 3-carboxy-2-hydroxyadipate - 644543 1.1.1.87 1.4 - homoisocitric acid - 644541 1.1.1.87 1.5 - alpha-ketoadipate - 644539 1.1.1.87 1.5 - homoisocitrate pH 7.8, 25°C, recombinant mutant R87T 660918 1.1.1.87 1.5 - NAD+ mutant enzyme T71V, with homoisocitrate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 1.5 - NAD+ wild type enzyme, with 3-isopropylmalate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 1.8 - NAD+ mutant enzyme I82N, with isocitrate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 2.1 - (2R,3S)-3-(2-hydroxyethyl)malate pH 7.8, 36°C, recombinant enzyme 667934 1.1.1.87 2.2 - isocitrate mutant enzyme I82N, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 2.7 - (2R,3S)-3-(4-pentenyl)malic acid pH 7.8, 28°C, recombinant enzyme 667934 1.1.1.87 2.8 - (2R,3S)-3-allylmalic acid pH 7.8, 28°C, recombinant enzyme 667934 1.1.1.87 3.2 - homoisocitrate reductive carboxylation of alpha-ketoadipate 685232 1.1.1.87 3.4 - 3-vinylmalate pH 7.8, 36°C, recombinant enzyme 667934 1.1.1.87 4.5 - D-malate wild type enzyme, with NAD+ as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 7.5 - (1R,2S)-1-hydroxybutane-1,2,4-tricarboxylate pH 7.8, 60°C 722569 1.1.1.87 7.5 - 1-hydroxy-1,2,4-butanetricarboxylate purified enzyme, pH 8.0, 60°C 656127 1.1.1.87 7.6 - NADP+ wild type enzyme, with homoisocitrate as cosubstrate, at pH 8.0 and 60°C 760522 1.1.1.87 9 - NAD+ pH 8.0, 25°C, in absence of K+ 696220 1.1.1.87 9.4 - (2R,3S)-3-(4-pentenyl)malic acid pH 7.8, 36°C, recombinant enzyme 667934 1.1.1.87 16.3 - CO2 reductive carboxylation of alpha-ketoadipate 685232