Modification of the active site of aldose reductase by nitric oxide

A. Chandra, S. Srivastava, J. M. Petrash, A. Bhatnagar, Satish Srivastava

Research output: Contribution to journalArticle

Abstract

Aldose reductase, the first enzyme of the polyol pathway of glucose metabolism, has been implicated in the etiology of secondary diabetic complications. However, its in vivo mechanisms of regulation arc not known. In this study we show that nitric oxide (NO) donors, sodium nitroprusside (SNP), S-nitroso-N-acetylpencillamine (SNAP), and 3-morpholinosydonemine (SIN-1) cause a time- and concentration-dependent loss of catalytic activity of wild type recombinant aldose reductase. The enzyme activity was partially recovered by dithiothreitol and sodium horohydride. NADPH and NADP protected the enzyme from NO-induced inactivation. Since in a complete system with both substrates, NADPH and glyceraldehyde, the enzyme was not protected against inactivation by NO-donors, it is concluded that the enzyme is modified during the catalytic cycle. In contrast to the wild type enzyme and mutant forms C303S and C80S the mutant C298S was unaffected by NO donors, indicating selective modification of the enzyme at cys-298. These studies suggest that by specific modification of the active site thiol (cys-298), NO may be an endogenous regulator of the polyol pathway and cellular metabolism associated with aldose reductase.

Original languageEnglish (US)
JournalFASEB Journal
Volume11
Issue number9
StatePublished - 1997

Fingerprint

aldehyde reductase
Aldehyde Reductase
active sites
nitric oxide
Catalytic Domain
Nitric Oxide
Nitric Oxide Donors
Enzymes
enzymes
NADP
NADP (coenzyme)
polyols
Metabolism
inactivation
glyceraldehyde
Glyceraldehyde
mutants
metabolism
dithiothreitol
Dithiothreitol

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Chandra, A., Srivastava, S., Petrash, J. M., Bhatnagar, A., & Srivastava, S. (1997). Modification of the active site of aldose reductase by nitric oxide. FASEB Journal, 11(9).

Modification of the active site of aldose reductase by nitric oxide. / Chandra, A.; Srivastava, S.; Petrash, J. M.; Bhatnagar, A.; Srivastava, Satish.

In: FASEB Journal, Vol. 11, No. 9, 1997.

Research output: Contribution to journalArticle

Chandra, A, Srivastava, S, Petrash, JM, Bhatnagar, A & Srivastava, S 1997, 'Modification of the active site of aldose reductase by nitric oxide', FASEB Journal, vol. 11, no. 9.
Chandra A, Srivastava S, Petrash JM, Bhatnagar A, Srivastava S. Modification of the active site of aldose reductase by nitric oxide. FASEB Journal. 1997;11(9).
Chandra, A. ; Srivastava, S. ; Petrash, J. M. ; Bhatnagar, A. ; Srivastava, Satish. / Modification of the active site of aldose reductase by nitric oxide. In: FASEB Journal. 1997 ; Vol. 11, No. 9.
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