谷胱甘肽合成酶的改进检测方法：γ-谷氨酰半胱氨酸合成酶和谷胱甘肽合成酶。

Improved assay of the enzymes of glutathione synthesis: gamma-glutamylcysteine synthetase and glutathione synthetase.

作者信息

Beutler E, Gelbart T

出版信息

Clin Chim Acta. 1986 Jul 15;158(1):115-23. doi: 10.1016/0009-8981(86)90122-1.

Abstract

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.

摘要

已开发出估算红细胞γ-谷氨酰半胱氨酸合成酶和谷胱甘肽合成酶的新方法。使γ-32P ATP达到平衡，直到γ磷酸基团和β磷酸基团被均匀标记。测量在谷胱甘肽合成酶（GC-S）的底物谷氨酸和半胱氨酸存在下，或在谷胱甘肽合成酶（GSH-S）的底物γ-谷氨酰半胱氨酸和甘氨酸存在下释放的32Pi的量。这通过将磷钼酸盐络合物萃取到异丁醇-苯中来完成。这些方法与时间和溶血产物浓度呈线性关系。给出了正常值。