Inhibition of two mitochondrial enzymes by gold (III) halo complexes. Evidence for a binding mechanism.
Fumarase (EC 4.2.1.2) and mitochondrial L-malate dehydrogenase (EC 1.1.1.37) were both inhibited by NaAuCl4 and KAuBr4. The inhibition for both was measured as a function of gold complex concentration and aquation time, and the NaAuCl4 inhibition was also measured in the presence of 0.15 M NaCl. Regeneration of the enzyme activity after NaAuCl4 inhibition using L-cysteine, L-methionine and NaCN was also investigated. Sodium dodecyl sulfate (SDS) acrylamide gel electrophoresis and amino acid analysis was performed on the NaAuCl4 inhibited enzymes as well as on ribonuclease A (EC 3.1.26.2), lysozyme (EC 3.2.1.17) and liver alcohol dehydrogenase (EC 1.1.1.1). It was observed that the inhibition was proportional to the gold complex concentration but decreased markedly after aquation of the complex. In the presence of NaCl the initial rate of inactivation is essentially unaffected unless the complex has been aquated and then the initial rate is increased. Gel electrophoresis on gold complex-enzyme mixtures show polymerization for ribonuclease and lysozyme and amino acid analysis indicates that no oxidation has taken place. From these results, a binding mechanism is postulated for the inhibition of the dehydrogenases by direct displacement of a halide ligand, probably by two groups on the enzyme, at least one of which may be a sulfur containing acid.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001665:Binding Sites / Q000187:drug effects; D001965:Bromides / Q000494:pharmacology; D002712:Chlorides / Q000494:pharmacology; D005649:Fumarate Hydratase / Q000037:antagonists & inhibitors*; D006046:Gold / Q000494:pharmacology*; D017612:Gold Compounds; D066298:In Vitro Techniques; D008291:Malate Dehydrogenase / Q000037:antagonists & inhibitors*; D008928:Mitochondria / Q000201:enzymology*; D011485:Protein Binding; D013552:Swine
D000596:Amino Acids; D001965:Bromides; D002712:Chlorides; D017612:Gold Compounds; D006046:Gold; C024568:gold tetrachloride, acid; C037129:gold tetrabromide, acid; D008291:Malate Dehydrogenase; D005649:Fumarate Hydratase