Purification and characterization of sorbitol dehydrogenase from bovine brain.
Sorbitol dehydrogenase (EC 1.1.1.14) was isolated from bovine brain and purified 3,000-fold to apparent homogeneity, as judged by polyacrylamide gel electrophoresis. The purified enzyme had a specific activity of 36 units/mg of protein; a molecular weight of 39,000 for each of the four identical subunits and 155,000 for the intact enzyme were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel exclusion chromatography, respectively. The presence of one Zn2+ per subunit was confirmed by atom absorption spectroscopy; inactivation of the enzyme by metal-chelating agents points to the essential role that Zn2+ plays in the catalytically competent enzyme. The enzyme is also inactivated by thiol-blocking reagents; with respect to inactivation by sodium pyrophosphate, sorbitol dehydrogenase is different from closely related alcohol dehydrogenase.
Wiesinger
H
H
Physiologisch-Chemisches Institut, Universität, Tübingen, F.R.G.
Hamprecht
B
B
D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002417:Cattle; D002614:Chelating Agents; D002850:Chromatography, Gel; D011756:Diphosphates / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007064:L-Iditol 2-Dehydrogenase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism; D046911:Macromolecular Substances; D008970:Molecular Weight; D009243:NAD / Q000494:pharmacology; D013054:Spectrophotometry, Atomic; D013401:Sugar Alcohol Dehydrogenases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D015032:Zinc / Q000032:analysis
D002614:Chelating Agents; D011756:Diphosphates; D046911:Macromolecular Substances; D013439:Sulfhydryl Reagents; D009243:NAD; D013401:Sugar Alcohol Dehydrogenases; D007064:L-Iditol 2-Dehydrogenase; D000449:Aldehyde Reductase; D015032:Zinc; C003319:sodium pyrophosphate