PMID: 36150Apr 12, 1979

The role of cysteine residues in the catalytic activity of glycerol-3-phosphate dehydrogenase

Biochimica Et Biophysica Acta
R E Smith, R MacQuarrie

Abstract

Glycerol-3-phosphate dehydrogenase (sn-glycerol-3-phosphate:NAD+ 2-oxido-reductase, EC 1.1.1.8) has been shown to be sensitive to inhibition by iodoacetate. The reaction of the enzyme with iodoacetate, which appears to be a simple bimolecular process, is accompanied by a corresponding loss of enzyme activity. In addition to changes in activity, the alkylation reaction was monitored by the incorporation of radioactivity from iodo[2-14C]acetate, by changes in amino acid composition, and by changes in the content of free sulfhydryl groups. It is concluded that there are two cysteine residues in the native dimeric enzyme which are essential for enzymic activity. The rate of inactivation was relatively insensitive to the presence of various compounds with the exception of NADH which markedly inhibited the reaction. Kinetic and binding studies showed that the binding of NADH prevents alkylation and, conversely, alkylation prevents NADH binding. From the pH dependence of the alkylation reaction, the pKa of the essential sulfhydryl groups was calculated to be 8.5 and it is suggested that the binding of coenzyme is independent of the state of ionization of these groups.

References

Jun 1, 1995·The International Journal of Biochemistry & Cell Biology·T C KoekemoerW Oelofsen
Jan 24, 2014·Redox Biology·Ryan J MaillouxWilliam G Willmore
Dec 19, 2014·Molecular BioSystems·D W Bak, E Weerapana
Feb 6, 2014·Physical Chemistry Chemical Physics : PCCP·Srabanti GhoshSomobrata Acharya

Citations

Jul 1, 1973·Proceedings of the National Academy of Sciences of the United States of America·M J AdamsS S Taylor
May 1, 1970·Archives of Biochemistry and Biophysics·B M AndersonC N Wang
Feb 15, 1973·Journal of Molecular Biology·S A Bernhard, R A MacQuarrie
May 15, 1973·Journal of Molecular Biology·S T Rao, M G Rossmann
Aug 1, 1971·Archives of Biochemistry and Biophysics·T P FondyD B Rutherford
May 1, 1959·Archives of Biochemistry and Biophysics·G L ELLMAN
Jul 10, 1963·Biochemical and Biophysical Research Communications·T K LI, B L VALLEE

Related Concepts

Nadh Binding
Glycerin
Glycerol-3-Phosphate Dehydrogenase
Phosphate Measurement
NADH
Sulfhydryl Compounds
Chinchilla Rabbits
Enzyme Activity
Blood Enzyme Activity (Lab Test)
Cysteine

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