Mechanism of inactivation of glyceraldehyde-3-phosphate dehydrogenase in the presence of methylglyoxal

• Published in: Archives of biochemistry and biophysics Vol. 733; p. 109485
• Main Authors: Barinova, K.V., Serebryakova, M.V., Melnikova, A.K., Medvedeva, M.V., Muronetz, V.I., Schmalhausen, E.V.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.01.2023
• Subjects: Cysteine - metabolism; Diabetes; GAPDH; Glyceraldehyde-3-Phosphate Dehydrogenases - chemistry; Humans; Hydroimidazolone; Magnesium Oxide; Methylglyoxal; Neuroblastoma; Pyruvaldehyde; ROS; Superoxide; PBS; MG-H1; DTT; Superoxide; EDTA; Methylglyoxal; GA-3-P; Hydroimidazolone; MGO; SH-groups; ROS; Diabetes; GAPDH
• ISSN: 0003-9861; 1096-0384
• DOI: 10.1016/j.abb.2022.109485

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is known to be one of the targets of methylglyoxal (MGO), a metabolite of glycolysis that increased in diabetes. However, the mechanism of GAPDH inactivation in the presence of MGO is unclear. The purpose of the work was to study the reaction of GAPDH with MGO and to identify the products of the reaction. It was shown that incubation of recombinant human GAPDH with MGO leads to irreversible inactivation of the enzyme, which is accompanied by a decrease in SH-group content by approximately 3.3 per tetramer GAPDH. MALDI-TOF MS analysis showed that the modification of GAPDH with MGO results in the oxidation of the catalytic cysteine residues (Cys152) to form cysteine-sulfinic acid. In addition, 2 arginine residues (R80 and R234) were identified that react with MGO to form hydroimidazolones. Incubation of SH-SY5Y neuroblastoma cells with MGO resulted in the inactivation of GAPDH and inhibition of glycolysis. The mechanism of GAPDH oxidation in the presence of MGO suggests the participation of superoxide anion, which is formed during the reaction of amino groups with methylglyoxal. The role of GAPDH in protection against the damaging effect of ROS in cells in the case of inefficiency of MGO removal by the GSH-dependent glyoxalase system is discussed. [Display omitted] •Incubation of GAPDH with methylglyoxal results in irreversible oxidation of Cys152.•The catalytic residue Cys152 is oxidized to form cysteine-sulfinic acid.•Arginine residues R80 and R234 are modified to form hydroimidazolones (MG-H1).•The mechanism of Cys152 oxidation suggests involvement of superoxide anion.