PMID: 2510598Nov 15, 1989Paper

Functional cysteinyl residues in human placental aldose reductase

Archives of Biochemistry and Biophysics
S Q LiuS K Srivastava

Abstract

Incubation of human placental aldose reductase (EC 1.1.1.21) with the sulfhydryl oxidizing reagents 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) and N-ethylmaleimide (NEM) results in a biexponential loss of catalytic activity. Inactivation by DTNB or NEM is prevented by saturating concentrations of NADPH. ATP-ribose offers partial protection against inactivation by DTNB, whereas NADP, nicotinamide mononucleotide (NMN), and the substrates glyceraldehyde and glucose offer little or no protection. The inactivation by DTNB was reversed by dithiothreitol and partially by 2-mercaptoethanol but not by KCN. When the release of 2-nitro-5-mercaptobenzoic acid was measured, 3 mol of sulfhydryl residues was found to be modified per mole of the enzyme by DTNB. Correlation of the fractional activity remaining with the extent of modification by the statistical method of C.-L. Tsou (1962, Sci. Sin. 11, 1535-1558) indicates that of the three reactive residues, one reacts at a faster rate than the other two, and that two residues are essential for the catalytic activity of the enzyme. Labeling of the total sulfhydryl by [14C]NEM and quantification of DTNB-reactive residues in the enzyme denatured by 6 M urea indicates that a total of seven sulfhyd...Continue Reading

References

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Citations

Jan 1, 1994·Molecular Aspects of Medicine·G Desoye, E Shafrir
Aug 7, 1993·Biochimica Et Biophysica Acta·S Q LiuS K Srivastava
Jan 5, 1995·Biochimica Et Biophysica Acta·D A CarperS E Old
Apr 13, 1994·Biochimica Et Biophysica Acta·A BhatnagarS K Srivastava
Oct 13, 2006·Journal of the American Chemical Society·Aaron T FafarmanSteven G Boxer
Oct 1, 1992·Biochemical Medicine and Metabolic Biology·A Bhatnagar, S K Srivastava
Jun 22, 2005·Experimental Eye Research·Seema SrivastavaSatish K Srivastava
Mar 4, 2014·Insect Molecular Biology·H GuoC-Z Wang
Jun 1, 1991·Archives of Biochemistry and Biophysics·A BhatnagarS K Srivastava
Dec 27, 1996·The Journal of Biological Chemistry·M CappielloU Mura
Mar 27, 2007·Journal of Pharmacological Sciences·Kazumi IwataChihiro Yabe-Nishimura
Dec 1, 1990·Archives of Biochemistry and Biophysics·A Del CorsoU Mura

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