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Effect of Gln on mitochondrial GSH, GSSG, S-glutathionylation, and complex I activity.

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posted on 2015-07-06, 06:16 authored by Kai Li, Yong-Chun Cui, Hong Zhang, Xiao-Peng Liu, Dong Zhang, Ai-Li Wu, Jian-Jun Li, Yue Tang

H9C2 cells were treated as described in Fig 3. (A) Mitochondrial GSH, (B) GSSG, and (C) the GSH/GSSG ratio in each group were analyzed. (D and E) S-glutathionylated Complex I protein expression (51—and 75—kDa subunits) from each group were detected by immunoprecipitation and western blot analysis. The up panel shows the anti-complex I 75 and 51 kDa subunits positive-IP sample followed by identification by an anti-GSH antibody. As a control, mitochondrial protein (20 μg) was also immunoblotted with complex I 75 kDa and 51 kDa subunit antibody. Bar graphs summarize the protein band intensities of S- glutathionylated Complex I. (F) Complex I activity (200 μg mitochondrial protein) of each group are detected. * P < 0.05 compare with the control group, # P < 0.05 compare with the 1 mM Gln and 4 mM Gln groups. The data are expressed as the mean ±S.D. of 3 independent experiments, a.u. = arbitrary units.

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