Monkey liver indanol dehydrogenase. Purification, properties, and kinetic mechanism.
Indanol dehydrogenase was purified to apparent homogeneity from monkey liver cytosol. The enzyme was a monomer with a molecular weight of 36,000 and pI of 8.7. The amino acid composition was determined. The enzyme oxidized alicyclic alcohols including transdihydrodiols of benzene and naphthalene in the presence of both NADP+ and NAD+, and reduced several xenobiotic carbonyl compounds in the presence of NADPH, the 4-pro-R hydrogen atom of which was transferred to the substrate. The results of fluorometric binding and kinetic studies are consistent with an ordered sequential mechanism with NADP+ binding first. The enzyme was inhibited competitively versus NADP+ and uncompetitively versus 1-indanol not only by chelating agents such as 1,10-phenanthroline and 2,2'-bipyridine but also by a nonchelating isomer, 4,4'-bipyridine, which suggests hydrophobic interaction of the aromatic compounds with the enzyme, which did not contain zinc. The enzyme was also inhibited by Cibacron blue dye, synthetic estrogens, and delta 4-3-ketosteroids. The inhibition by Cibacron blue was competitive versus NADP+ and noncompetitive versus 1-indanol, whereas those by hexestrol, medroxyprogesterone acetate, and progesterone were uncompetitive versus NADP+ and competitive versus 1-indanol, corraborating the ordered addition of the coenzyme prior to 1-indanol.
Hara
A
A
Department of Biochemistry, Gifu Pharmaceutical University.
Mouri
K
K
Nakagawa
M
M
Nakamura
M
M
Nakayama
T
T
Matsuura
K
K
Sawada
H
H
D000429:Alcohol Oxidoreductases / Q000032:analysis / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002614:Chelating Agents; D055598:Chemical Phenomena; D002621:Chemistry; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008251:Macaca; D008297:Male; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D013379:Substrate Specificity; D014018:Tissue Distribution
D000596:Amino Acids; D002614:Chelating Agents; D009243:NAD; D009249:NADP; D000429:Alcohol Oxidoreductases; C046532:indanol dehydrogenase