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

Natalie Z.M. Homer, J. Reglinski, Rebecca J. Sowden, C.M. Spickett, Rhoda Wilson, James J. Walker

Research output: Contribution to journalArticle

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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.
LanguageEnglish
Pages317-324
Number of pages7
JournalCryobiology
Volume50
Issue number3
DOIs
Publication statusPublished - Jun 2005

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dimethyl sulfoxide
Dimethyl Sulfoxide
Glutathione
glutathione
erythrocytes
Erythrocytes
Nuclear magnetic resonance
kinetics
Kinetics
oxidation
Oxidation
Glutathione Reductase
Cryopreservation
glutathione-disulfide reductase
thiols
Sulfhydryl Compounds
cryopreservation
In Vitro Techniques
Rate constants
Cells

Keywords

  • cryoprotectant
  • dimethylsulfoxide
  • glutathione
  • erythrocyte
  • H-1 NMR
  • oxidative stress

Cite this

Homer, N. Z. M., Reglinski, J., Sowden, R. J., Spickett, C. M., Wilson, R., & Walker, J. J. (2005). Dimethylsulfoxide oxidizes glutathione in vitro and in human erythrocytes: kinetic analysis by H-1 NMR. Cryobiology, 50(3), 317-324. https://doi.org/10.1016/j.cryobiol.2005.04.002
Homer, Natalie Z.M. ; Reglinski, J. ; Sowden, Rebecca J. ; Spickett, C.M. ; Wilson, Rhoda ; Walker, James J. / Dimethylsulfoxide oxidizes glutathione in vitro and in human erythrocytes: kinetic analysis by H-1 NMR. In: Cryobiology. 2005 ; Vol. 50, No. 3. pp. 317-324.
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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.",
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Homer, NZM, Reglinski, J, Sowden, RJ, Spickett, CM, Wilson, R & Walker, JJ 2005, 'Dimethylsulfoxide oxidizes glutathione in vitro and in human erythrocytes: kinetic analysis by H-1 NMR' Cryobiology, vol. 50, no. 3, pp. 317-324. https://doi.org/10.1016/j.cryobiol.2005.04.002

Dimethylsulfoxide oxidizes glutathione in vitro and in human erythrocytes: kinetic analysis by H-1 NMR. / Homer, Natalie Z.M.; Reglinski, J.; Sowden, Rebecca J.; Spickett, C.M.; Wilson, Rhoda; Walker, James J.

In: Cryobiology, Vol. 50, No. 3, 06.2005, p. 317-324.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Dimethylsulfoxide oxidizes glutathione in vitro and in human erythrocytes: kinetic analysis by H-1 NMR

AU - Homer, Natalie Z.M.

AU - Reglinski, J.

AU - Sowden, Rebecca J.

AU - Spickett, C.M.

AU - Wilson, Rhoda

AU - Walker, James J.

PY - 2005/6

Y1 - 2005/6

N2 - 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.

AB - 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.

KW - cryoprotectant

KW - dimethylsulfoxide

KW - glutathione

KW - erythrocyte

KW - H-1 NMR

KW - oxidative stress

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