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Dimethylsulfoxide oxidizes glutathione in vitro and in human erythrocytes: kinetic analysis by H-1 NMR

Homer, Natalie Z.M. and Reglinski, J. and Sowden, Rebecca J. and Spickett, C.M. and Wilson, Rhoda and Walker, James J. (2005) Dimethylsulfoxide oxidizes glutathione in vitro and in human erythrocytes: kinetic analysis by H-1 NMR. Cryobiology, 50 (3). pp. 317-324. ISSN 0011-2240

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Abstract

The interaction of dimethylsulfoxide (Me2SO) with glutathione was investigated under non-equilibrium conditions in solution using 1H NMR and in intact erythrocytes using 1H spin-echo NMR. In solution the reaction was observed to follow second-order kinetics (Rate = k1[glutathione][Me2SO]) at 300 K pH 7.4, ksol = 4.7 × 10−5 mol −1 L1 s−1. In intact erythrocytes the rate constant for the cellular environment, kcell, was found to be slightly larger at 8.1 × 10−5 mol−1 L1 s−1. Furthermore, the reaction of Me2SO with erythrocyte glutathione showed a biphasic dependence on the Me2SO concentration, with little oxidation of glutathione occurring until the Me2SO concentration exceeded 0.5 mol L−1. The results suggest that at lower concentrations, Me2SO can be effectively removed, most probably by reaction with glutathione, which is regenerated by glutathione reductase, although preferential reaction with other cellular components (e.g., membrane or cellular thiols) cannot be ruled out. Thus the concentrations of Me2SO that are commonly used in cryopreservation of mammalian cells (1.4 mol L−1) can cause oxidation of intracellular glutathione.

Item type: Article
ID code: 17284
Keywords: cryoprotectant, dimethylsulfoxide, glutathione, erythrocyte, H-1 NMR, oxidative stress, Pharmacy and materia medica, Agricultural and Biological Sciences(all), Biochemistry, Genetics and Molecular Biology(all), Medicine(all)
Subjects: Medicine > Pharmacy and materia medica
Department: Faculty of Science > Strathclyde Institute of Pharmacy and Biomedical Sciences
Faculty of Science > Pure and Applied Chemistry
Related URLs:
    Depositing user: Strathprints Administrator
    Date Deposited: 13 Apr 2010 15:21
    Last modified: 05 Sep 2014 01:26
    URI: http://strathprints.strath.ac.uk/id/eprint/17284

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