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J. Biochem, 1990, Vol. 108, No. 1 9-16
© 1990 Japanese Biochemical Society

research-article

Kinetics of Hydroperoxide Degradation by NADP-Glutathione System in Mitochondria1

Kazuhei Kurosawa*, Hideo Shibata*, Norio Hayashi**, Nobuhiro Sato**, Takenobu Kamada** and Kunio Tagawa*,2

*Department of Physiological Chemistry Kita-ku, Osaka, Osaka 530
**First Department of Medicine, Osaka University Medical School Kita-ku, Osaka, Osaka 530

2To whom correspondence should be addressed

Hydroperoxide decomposition by the NADP-glutathione system in rat liver mitochondria was analyzed. Mitochondria were found to contain high concentrations of the reduced form of glutathione (GSH) (4.32±0.50nmol/mg) and NADPH (4.74±0.64nmol/mg), and high activities of glutathione peroxidase and reductase. In the initial phase of the reaction, the rate of hydroperoxide decomposition was proportional to both the GSH level and the activity of GSH peroxidase. However, in the later steady state, the step of NADP reduction was rate-limiting, and the overall reaction rate was independent of the initial concentration of GSH, and activities of glutathione peroxidase and reductase. Some GSH was released from mitochondria during incubation, but the rate of the decomposition could be simply expressed as k [GSH]/2, where k is the first-order rate constant of the peroxidase and [GSH] is the intramitochondrial level of GSH in the steady state. The rate of the reaction in the steady state was also dependent on the NADPH level, its reciprocal being linearly correlated with [NADPH]–1. The rate of decomposition of hydroperoxide was influenced by the respiratory state. During state 3 respiration, the rate was greatly depressed, but was still considered to exceed by far the rate of physiological generation of hydroperoxide.

1This study was supported by a Grant-in-Aid for Scientific Research (A63440025) from the Ministry of Education, Science and Culture of Japan, and by a grant from the Nisshin Seifun Foundation.


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