Turnover of Cellular Glutathione in Isolated Rat‐Kidney Cells

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Abstract
Turnover of cellular glutathione in isolated rat kidney cells was studied using cystine or Met as S donor. In the absence of S donor a continuous decrease of intracellular reduced glutathione (GSH) during incubation of the cells was observed. This decrease was abolished in the presence of cystine and, as indicated by incorporation of 35S, there was also a rapid synthesis of GSH. In the presence of a .gamma.-glutamyltransferase inhibitor, the synthesis of intracellular GSH was accompanied by an accumulation of extracellular cysteine-glutathione mixed disulfide whereas only minor amounts of GSH and glutathione disulfide could be detected. The intracellular levels of both the cystine-glutathione and glutathione disulfides were at all time points very low. Even though the uptake of cystine was rapid and not rate-limiting for GSH synthesis, almost no cystine could be detected intracellularly. An increasing intracellular cysteine concentration was observed, indicating a rapid reduction of cystine. In contrast to cystine, Met did not protect from the loss of intracellular GSH and only a low rate of incorporation of 35S into GSH was observed. Met was rapidly taken up into the cells but was apparently converted to cysteine only to a very limited extent. This is most likely due to a low activity of the enzyme cystathionase since neither homocysteine nor cystathionine was very effective in supporting GSH synthesis.