Effect of Resveratrol and Tiron on the Inactivation of Glyceraldehyde-3- phosphate Dehydrogenase Induced by Superoxide Anion Radical

Abstract

Protein damage mediated by oxidation has been associated with aging and age-related diseases, in particular neurodegenerative diseases. The protein that is known to be one of the major targets of oxidative stress is glyceraldehyde- 3-phosphate dehydrogenase. GAPDH is believed to play a key role in certain neurodegenerative disorders, such as Alzheimer's, Parkinson's, and Huntington's diseases. Several recent studies have suggested that a wide range of variety of polyphenols including resveratrol may have neuroprotective effects. Here, we present data that clearly indicate the prooxidative properties of resveratrol and tiron in the inactivation of GAPDH induced by the superoxide anion generated via xanthine oxidase mediated oxidation of xanthine. Generated in the studied system tiron and resveratrol radicals are much more efficient in the inactivation of GAPDH than the superoxide anion alone. The analysis of CD spectra of protein exposed to the tiron and resveratrol radicals revealed little effect on the secondary structure of GAPDH. In both cases reduction of α-helical structure was followed by the increase in β-sheet conformation. Thus, the most probable mechanism of inactivation of GAPDH in the studied system is oxidation of cysteine residues in the catalytic center of the enzyme. Finally, molecular modeling of the resveratrol - GAPDH and tiron - GAPDH complexes showed potential binding sites for those antioxidants with binding affinity -45 kcal/mol and -48 kcal/mol respectively.

Comparison of Antioxidants: The Limited Correlation between Various Assays of Antioxidant Activity

“…We and others have shown such that other antioxidants, melatonin and resveratrol showed prooxidant effects on glyceraldehyde 3-phosphate dehydrogenase when applied together with nitric oxide; this effect was due to formation of their reactive derivatives, viz. phenoxyl radical in the case of resveratrol [ 36 , 37 ]. As a phenoxyl radical is formed also during oxidation of gentisic acid [ 30 ], the mechanism of the prooxidant action of gentisic acid may be similar.…”

Determination of trans-resveratrol action on two different types of neuronal cells, neuroblastoma and hippocampal cells

“…The percentage of apoptotic cells was higher than what would be expected from the combined action of the two factors -hydrogen peroxide and resveratrol. Intensification of apoptotic processes in cancer cells may be the result of, among other influences, oxidative damage to different biomolecules caused by the formation of phenoxyl radicals of resveratrol, efflux of cytochrome C, and the influence of RSV on mitochondrial membrane proteins, which may result in a decrease in the mitochondrial potential (van Ginkel et al 2007, Li et al 2012Xi et al 2012;Rodacka et al 2014). The greater sensitivity of tumour cells to the action of resveratrol or its metabolites in combination with hydrogen peroxide may also be due to conditions in the tumour microenvironment -low pH, reduced activity of antioxidant enzymes, increased oxidative stress (Shamim et al 2012).…”

Glyceraldehyde-3-phosphate Dehydrogenase Is a Multifaceted Therapeutic Target

“…The most probable site for the oxidation is the C152 residue located in the catalytic region of the enzyme molecule. Oxidation of C152 causes an inactivation of GAPDH and the formation of intra-or inter-molecular disulfide bonds [64,65]. Another mechanism triggered by the oxidation of M46 on the GAPDH molecule, which also leads to enzyme denaturation and the formation of aggregates through disulfide bridges [66].…”