Cellular recovery of glyceraldehyde-3-phosphate dehydrogenase activity and thiol status after exposure to hydroperoxides

doi:10.1016/0003-9861(90)90028-W

Archives of Biochemistry and Biophysics

Volume 276, Issue 1, January 1990, Pages 212-218

https://doi.org/10.1016/0003-9861(90)90028-W Get rights and content

Abstract

The activity of the thiol-dependent enzyme glyceraldehyde-3-phosphate dehydrogenase (GPD), in vertebrate cells, was modulated by a change in the intracellular thiol:disulfide redox status. Human lung carcinoma cells (A549) were incubated with 1–120 mm H₂O₂, 1–120 mm t-butyl hydroperoxide, 1–6 mm ethacrynic acid, or 0.1–10 mmN-ethylmaleimide for 5 min. Loss of reduced protein thiols, as measured by binding of the thiol reagent iodoacetic acid to GPD, and loss of GPD enzymatic activity occurred in a dose-dependent manner. Incubation of the cells, following oxidative treatment, in saline for 30 min or with 20 mm dithiothreitol (DTT) partially reversed both changes in GPD. The enzymatic recovery of GPD activity was observed either without addition of thiols to the medium or by incubation of a sonicated cell mixture with 2 mm cysteine, cystine, cysteamine, or glutathione (GSH); GSSG had no effect. Treatment of cells with buthionine sulfoximine (BSO) to decrease cellular GSH by varying amounts caused a dose-related increase in sensitivity of GPD activity to inactivation by H₂O₂ and decreased cellular ability for subsequent recovery. GPD responded in a similar fashion with oxidative treatment of another lung carcinoma cell line (A427) as well as normal lung tissue from human and rat. These findings indicate that the cellular thiol redox status can be important in determining GPD enzymatic activity.

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Cellular recovery of glyceraldehyde-3-phosphate dehydrogenase activity and thiol status after exposure to hydroperoxides

Abstract

Biochim. Biophys. Acta

Int. J. Biochem

Free Rad. Biol. Med

Biochem. Biophys. Res. Commun

Anal. Biochem

Biochim. Biophys. Acta

J. Biol. Chem

Anal. Biochem

J. Biol. Chem

Toxicol. Appl. Pharmacol

J. Biol. Chem

J. Biol. Chem

Exp. Eye Res