Rat lens aldose reductase: rapid purification and comparison with human placental aldose reductase

Roland K. Herrmann, Peter F. Kador, Jin H. Kinoshita

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

Abstract

Aldose reductase (alditol: NADP oxidoreductase EC.1.1.1.21), an enzyme in the sorbitol pathway which has been implicated in the pathogenesis of diabetic complications, has been purified from rat lens (RLAR) by affinity chromatography with Amicon Matrex Gel Orange A and its properties have been compared to those of purified human placental aldose reductase (HPAR). The RLAR appears to be closely associated with α- and β-crystallin and has a higher affinity for the dye Matrex column than HPAR. The purified enzyme, obtained upon elution from the column, appears as a closely-spaced doublet of approximately 38 K MW on SDS-PAGE which does not immunologically cross-react with antibodies raised against the single 38 K MW HPAR. Antibodies raised against RLAR however do cross-react with HPAR. Kinetic studies indicated both enzymes to have a greater apparent affinity for aliphatic and aromatic aldehydes than for aldose sugars. Compared to dl-glyceraldehyde, RLAR displayed an 80-fold greater affinity for p-nitrobenzaldehyde and a 1000-fold decreased affinity for d-glucose while HPAR displayed a 15-fold greater affinity for p-nitrobenzaldehyde and 600-fold less affinity for d-glucose. Both enzymes displayed only trace activity with 200 mm l-gulonic acid.

Original languageEnglish (US)
Pages (from-to)467-474
Number of pages8
JournalExperimental Eye Research
Volume37
Issue number5
DOIs
StatePublished - Nov 1983
Externally publishedYes

Keywords

  • aldose reductase
  • diabetic complications
  • purification
  • rat lens

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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