Oxygen dependence of glutathione synthesis in hepatocytes

The O2 dependence of glutathione (GSH) synthesis was studied in freshly isolated hepatocytes of white male rats. The rate of synthesis with methionine as the sulfur-containing amino acid precursor was decreased at hypoxic O2 concentrations and was half-maximal at 5 microM O2. ATP-dependent formation of S-adenosylmethionine was the rate-limiting step in GSH synthesis under these hypoxic conditions as shown by studies of S-adenosylmethionine concentrations and effects of compounds that inhibit mitochondrial ATP production. GSH synthesis with cysteine as the sulfur-containing precursor amino acid was relatively resistant to O2 deficiency. The rate under anoxia was 48% of the aerobic rate and the O2-dependent rate was half-maximal at 0.9 microM O2. These results indicate that GSH synthesis from methionine is likely to be impaired under physiological and pathological conditions involving hypoxia, but synthesis from cysteine is not likely to be greatly affected except during anoxia. In addition, the sensitivity of the cystathionine pathway to hypoxia suggests that other products of the pathway, such as choline, creatine, epinephrine, and methylated tRNA's, may also be decreased by hypoxia.