Partial catalytic Cys oxidation of human GAPDH to Cys-sulfonic acid.

Abstract

Background: n-Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) catalyses the NAD+-dependent oxidative phosphorylation of n-glyceraldehyde-3-phosphate to 1,3-diphospho-n-glycerate and its reverse reaction in glycolysis and gluconeogenesis. Methods: Four distinct crystal structures of human n-Glyceraldehyde-3-phosphate dehydrogenase (HsGAPDH) have been determined from protein purified from the supernatant of HEK293F human epithelial kidney cells. Results: X-ray crystallography and mass-spectrometry indicate that the catalytic cysteine of the protein (HsGAPDH Cys152) is partially oxidised to cysteine S-sulfonic acid. The average occupancy for the Cys152-S-sulfonic acid modification over the 20 crystallographically independent copies of HsGAPDH across three of the crystal forms obtained is 0.31±0.17. Conclusions: The modification induces no significant structural changes on the tetrameric enzyme, and only makes aspecific contacts to surface residues in the active site, in keeping with the hypothesis that the oxidising conditions of the secreted mammalian cell expression system result in HsGAPDH catalytic cysteine S-sulfonic acid modification and irreversible inactivation of the enzyme.

Reactive Sulphur Species and Exposome: A Perspective on Potential Role in Alleviating UV-Induced Stress

“…2). Over-oxidation of the protein thiol group, which leads to the formation of sulfinic acid (RSO 2 H) and sulfonic acid (RSO 3 H) has been implicated with irreversible post-translational modification [43][44][45][46][47][48]. Such modification can render the enzymes or proteins to become dysfunctional.…”

Redox–Oligomeric State of Peroxiredoxin-2 and Glyceraldehyde-3-Phosphate Dehydrogenase in Obstructive Sleep Apnea Red Blood Cells under Positive Airway Pressure Therapy

“…However, once sulfonylated GAPDH is formed, it is generally considered as an irreversible inactive form associated with protein misfolding prone to degradation [48]. Very recently, Lia et al, by determining the first crystal structure of GAPDH tetramer carrying sulfonylated modification in active Cys, showed that this modification induces an irreversible inactivation of the enzyme without causing significant structural changes on the tetrameric enzyme [49]. However, the exact "gain of function" associated with this overoxidation remains to be elucidated.…”

Stoichiometric Thiol Redox Proteomics for Quantifying Cellular Responses to Perturbations

“…As an illustration, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), an important enzyme in the glycolytic pathway, is susceptible to multiple types of thiol redox modifications, such as nitrosylation at the active site residue Cys150 of mouse GAPDH [ 25 ]. A more recent study has found that the corresponding cysteine in human GAPDH, Cys152, is subject to sulfonic acid formation that irreversibly inactivates GAPDH [ 26 ]. Sulfenylation at the same residue in rabbit GAPDH in response to hydrogen peroxide exposure has also been reported [ 27 ], where the sulfenic acid promotes a reaction with reduced glutathione.…”