Regulation of glutamate dehydrogenase by Mg2+ and magnification of leucine activation by Mg2+.
In the presence of Mg2+, pure glutamate dehydrogenase is more reactive with NADPH than with NADH and is markedly activated by elevations in the ADP/ATP ratio or the addition of leucine. Because these are properties of glutamate dehydrogenase in mitochondria but not properties of the pure enzyme studied in the absence of Mg2+, Mg2+ could be a ligand that confers upon glutamate dehydrogenase the regulatory properties of this enzyme found in situ. In the absence of the allosteric activators ADP, leucine, or succinyl-CoA, Mg2+ is an inhibitor and increases product inhibition by alpha-ketoglutarate in the forward reaction and substrate inhibition by alpha-ketoglutarate in the reverse reaction. However, the allosteric activators convert Mg2+ from an inhibitor into an activator of the forward reaction. In the reverse reaction, ADP also converts Mg2+ from an inhibitor into an activator and leucine eliminates inhibition by Mg2+. Because Mg2+ is an inhibitor in the absence of activator that also increases inhibition by alpha-ketoglutarate, whereas in the presence of activator Mg2+ has no effect or is itself an activator, Mg2+ magnifies the effect of the activator, and magnification increases with increases in the concentration of alpha-ketoglutarate. Leucine and its analog 2-aminobicyclo (2.2.1) heptane 2-carboxylic acid (BCH) have almost identical effects on both human and bovine glutamate dehydrogenase in both the presence and absence of Mg2+. However, advantages of BCH over leucine as a potential pharmacological activator of glutamate dehydrogenase are that BCH is not metabolized and, unlike leucine, BCH does not inhibit ornithine transcarbamylase. Isoleucine and valine alone have little effect on human glutamate dehydrogenase, but isoleucine slightly inhibits the enzyme in the presence of leucine.
Fahien
L A
LA
Department of Pharmacology, University of Wisconsin Medical School, Madison 53706.
Teller
J K
JK
Macdonald
M J
MJ
Fahien
C M
CM
D000494:Allosteric Regulation; D000596:Amino Acids / Q000494:pharmacology; D000598:Amino Acids, Cyclic; D004789:Enzyme Activation; D005969:Glutamate Dehydrogenase / Q000378:metabolism*; D006801:Humans; D007656:Ketoglutaric Acids / Q000494:pharmacology; D007700:Kinetics; D007930:Leucine / Q000494:pharmacology*; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008274:Magnesium / Q000494:pharmacology*; D008297:Male; D009954:Ornithine Carbamoyltransferase / Q000378:metabolism
D000596:Amino Acids; D000598:Amino Acids, Cyclic; D007656:Ketoglutaric Acids; C006504:2-aminobicyclo(2,2,1)heptane-2-carboxylic acid; D005969:Glutamate Dehydrogenase; D009954:Ornithine Carbamoyltransferase; D007930:Leucine; D008274:Magnesium