Competition between inhibitors of the trypanosome alternative oxidase (TAO) and reduced coenzyme Q9.
The trypanosome alternative oxidase (TAO) is an attractive target for chemotherapy for the diseases caused by African trypanosomes because there is no equivalent enzyme in mammalian hosts. Many inhibitors of this enzyme have been described, but there have been no data on the mechanism of inhibition. In the present study, reduced 2,3-dimethoxy-5-methyl-6-decyl-1,4-benzoquinone (decyl-CoQ-H2) was used as a substitute for the natural substrate CoQ9-H2 to allow direct measurements of the TAO in crude mitochondrial preparations from Trypanosoma brucei brucei. A Km value of 3.8 microM was obtained for this substrate. The following five compounds that have alkyl side chains from 1 to 4 carbons and belong to three classes of inhibitors showed a competitive inhibition pattern with respect to decyl-CoQ-H: p-methoxybenzhydroxamic acid, p-ethoxybenzhydroxamic acid,p-n-butyloxybenzhydroxamic acid, methyl 3,4-dihydroxybenzoate and N-n-butyl-3,4-dihydroxybenzamide. The following four compounds belonging to the same chemical classes but having alkyl side chains from 10 to 12 carbons showed uncompetitive inhibition patterns: p-n-dodecyloxybenzhydroxamic acid, n-decyl 3,4-dihydroxybenzoate, n-dodecyl 3,4-dihydroxybenzoate, and N-n-decyl-3,4-dihydroxybenzamide. Clearly, the first group of inhibitors compete with CoQ-H2 for the active site of the TAO. We propose that the uncompetitive patterns produced by the second group of inhibitors are due to the greater lipophilicity of these compounds and the resulting change in the interaction of the inhibitors and the membrane containing the TAO, thus affecting the local concentration of the inhibitors at the active site.
D000818:Animals; D000977:Antiparasitic Agents; D001665:Binding Sites; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005260:Female; D007700:Kinetics; D024101:Mitochondrial Proteins; D010088:Oxidoreductases / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D010940:Plant Proteins; D051381:Rats; D017207:Rats, Sprague-Dawley; D014344:Trypanocidal Agents / Q000494:pharmacology*; D014346:Trypanosoma brucei brucei / Q000201:enzymology*; D014451:Ubiquinone / Q000031:analogs & derivatives / Q000037:antagonists & inhibitors* / Q000138:chemical synthesis; D000099188:Alternative Oxidase
D000977:Antiparasitic Agents; D004791:Enzyme Inhibitors; D024101:Mitochondrial Proteins; D010088:Oxidoreductases; D010940:Plant Proteins; D014344:Trypanocidal Agents; D014451:Ubiquinone; D000099188:Alternative Oxidase; C060262:2,3-dimethoxy-5-methyl-6-decyl-1,4-benzoquinone