Inhibitory effect of vanadium compounds on glutamate dehydrogenase activity in mitochondria and hepatocytes isolated from rabbit liver.

Abstract

The effect of orthovanadate, vanadyl sulphate and vanadyl acetylacetonate on glutamate dehydrogenase activity was studied in liver mitochondria and isolated hepatocytes of rabbit. In permeabilized mitochondria with free access of substrates and drugs to glutamate dehydrogenase, orthovanadate and vanadyl sulphate at 200 microM concentrations decreased both glutamate synthesis and glutamate deamination by 80 and 50%, respectively, while vanadyl acetylacetonate was less potent. In view of kinetic data obtained at various ammonium concentrations, orthovanadate appeared to be a competitive inhibitor (Ki = 40 +/- 3 microM), while vanadyl sulphate was a non-competitive one (Ki = 147 +/- 10 microM). In contrast to orthovanadate, vanadyl sulphate augmented the inhibitory action of increased above 0.5 mM 2-oxoglutarate concentrations. All these effects on the enzyme activity were partially reversed in the presence of L-leucine and ADP, which are allosteric activators of glutamate dehydrogenase. Moreover, all compounds studied suppressed both glutamate formation and glutamate deamination in isolated hepatocytes incubated under various metabolic conditions, as concluded from decreased rates of glutamate and urea synthesis, respectively. In view of these observations it seems likely that vanadium-containing compounds may be potent inhibitors of glutamate metabolism in liver.