Bovine lens aldehyde reductase (aldose reductase). Purification, kinetics and mechanism

Biochem J. 1984 Apr 1;219(1):33-9. doi: 10.1042/bj2190033.

Abstract

Aldehyde reductase (aldose reductase) was purified to homogeneity (as judged by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis) from bovine lens by affinity chromatography on NADP+-Sepharose. The enzyme, a monomer of Mr about 40000, was active with a variety of alpha- hydroxyketones , including fructose. The minimum degree of the rate equation was 2:2 in the case of DL-glyceraldehyde, but linear kinetics were observed for glucose and NADPH over the concentration range studied. The enzyme largely followed a ternary-complex mechanism, with initial binding of NADPH before glucose and final release of NADP+.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Aldehyde Reductase* / antagonists & inhibitors
  • Aldehyde Reductase* / isolation & purification
  • Aldehyde Reductase* / metabolism
  • Amino Acids / analysis
  • Animals
  • Cattle
  • Chromatography, Affinity
  • Glucose / metabolism
  • Glyceraldehyde / metabolism
  • Kinetics
  • Lens, Crystalline / enzymology*
  • Molecular Weight
  • NADP / metabolism
  • Osmolar Concentration
  • Substrate Specificity
  • Sugar Alcohol Dehydrogenases* / isolation & purification
  • Sugar Alcohol Dehydrogenases* / metabolism

Substances

  • Amino Acids
  • Glyceraldehyde
  • NADP
  • Sugar Alcohol Dehydrogenases
  • Aldehyde Reductase
  • Glucose