Inhibition of alcohol dehydrogenase from yeast by pyridine

• Published in: Biochemical journal 1967-09, Vol.104 (3), p.872-877
• Main Authors: Atkinson, M R, Eckermann, G, Lilley, R M
• Format: Article
• Language: English
• Subjects: Alcohol Oxidoreductases - antagonists & inhibitors; Binding Sites; Chemical Phenomena; Chemistry, Physical; Ethanol; Kinetics; NAD; Pyridines - pharmacology; Yeasts - enzymology; Zinc
• ISSN: 0264-6021; 1470-8728
• DOI: 10.1042/bj1040872

1. Inhibition by pyridine of reduction of NAD by ethanol in the presence of yeast alcohol dehydrogenase was studied at 25 degrees in 60mm-glycine buffer (K(+), pH9.3). 2. The apparent Michaelis constant for ethanol increases linearly and that for NAD increases non-linearly with pyridine concentration. 3. Rates, v, observed in the presence of pyridine are lower than the values calculated from the effect of pyridine on the two apparent Michaelis constants and are described by the expression V/v={1+5.8[pyridine]}x{1+0.016(1+124 [pyridine)]/[EtOH]}x{1+0.00019(1+3.3[pyridine]+110 [pyridine](2))/[NAD]}. 4. Mixed inhibitor studies with pyridine and N(1)-methylnicotinamide chloride in 40mm-pyrophosphate buffer (Na(+), pH8.2) indicated little interaction of pyridine with the ;pyridinium site' of the dehydrogenase (interaction constant, alpha, 2.1). 5. The possible competition of ethanol and pyridine for a zinc atom in the active centre of yeast alcohol dehydrogenase is discussed.