Improved assay of the enzymes of glutathione synthesis: gamma-glutamylcysteine synthetase and glutathione synthetase.
New methods for the estimation of red cell gamma-glutamylcysteine synthetase and glutathione synthetase have been developed. gamma-32P ATP is allowed to equilibrate until the gamma and beta phosphate groups are equally labelled. The amount of 32Pi released in the presence of glutamic acid and cysteine, the substrates for GC-S or in the presence of gamma-glutamylcysteine and glycine, the substrates of GSH-S, is measured. This is accomplished by extraction of the phosphomolybdate complex into isobutanol-benzene. The methods are linear with time and hemolysate concentration. Normal values are presented.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002642:Chick Embryo; D004912:Erythrocytes / Q000201:enzymology; D005260:Female; D005721:Glutamate-Cysteine Ligase / Q000097:blood*; D005981:Glutathione Synthase / Q000097:blood* / Q000172:deficiency; D006801:Humans; D008297:Male; D010453:Peptide Synthases / Q000097:blood*; D010761:Phosphorus Radioisotopes; D012016:Reference Values
D010761:Phosphorus Radioisotopes; D000255:Adenosine Triphosphate; D010453:Peptide Synthases; D005721:Glutamate-Cysteine Ligase; D005981:Glutathione Synthase