Crystallization and partial characterization of glutamate dehydrogenase from ox liver nuclei.
Glutamate dehydrogenase have been obtained in crystalline form from purified ox liver nuclear fractions. The enzyme appeared homogeneous, as judged by several electrophoretic techniques at two pH values. A comparative study with the widely known ox liver mitochondrial glutamate dehydrogenase revealed several common features, such as the allosteric effect of the nucleotides ADP and GTP, the activation at high concentrations of the cofactor NAD+, and the existence of a concentration-dependent reversible monomer-polymer(s) equilibrium. However, the two enzymes differed in many other respects. Inorganic phosphate activated nuclear glutamate dehydrogenase to a much greater extent than the mitochondrial enzyme; the substrate NH4+ showed cooperative homotropic interactions only with nuclear glutamate dehydrogenase; kinetic differences were detected with most of the reaction substrates, as well as different rates of oxidative deamination of other L-amino acids, the nuclear enzyme had a higher anodic mobility and a different chromatographic behavior on anionic exchangers. The latter evidence indicates that the glutamate dehydrogenase activity in liver is associated with two proteins which are structurally different, thus confirming the results of a separate immunological study. Preliminary evidence suggests that the enzyme in nuclei is attached to the nuclear envelope, probably the inner membrane, from which it can be solubilized by the addition of salts.
D000494:Allosteric Regulation; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000201:enzymology*; D003460:Crystallization; D003641:Deamination; D004789:Enzyme Activation; D005969:Glutamate Dehydrogenase / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000201:enzymology*; D010084:Oxidation-Reduction; D012265:Ribonucleotides / Q000494:pharmacology
D000596:Amino Acids; D012265:Ribonucleotides; D005969:Glutamate Dehydrogenase