谷胱甘肽 - 前列腺素A1共轭物作为从兔肾中纯化前列腺素9 - 酮还原酶的底物。

Glutathione-prostaglandin A1 conjugate as substrate in the purification of prostaglandin 9-ketoreductase from rabbit kidney.

作者信息

Toft B S, Hansen H S

出版信息

Prostaglandins. 1980 Oct;20(4):735-46. doi: 10.1016/0090-6980(80)90112-4.

Abstract

Using GSH-PGA1 as substrate for determination of enzyme activity a pI 4.8 form of rabbit kidney prostaglandin 9-keto-reductase has been purified 95 times to a specific activity of 1755 nmol/min per mg protein. The purification procedures involve ion-exchange chromatography, gel-filtration and affinity chromatography. The latter procedure comprises Blue Sepharose affinity chromatography, and GSH-PGA1-Sepharose affinity chromatography. The purified enzyme preparation also showed a weak NADP+-dependent 15-hydroxyprostaglandin dehydrogenase activity, 20 nmol/min per mg protein with PGE1 as substrate. Km (PGE1) for the dehydrogenase is 142.6 +/- 45.1 microM (S.E., n = 7).

摘要

以谷胱甘肽 - PGA1 为底物测定酶活性时，兔肾前列腺素 9 - 酮还原酶的一种 pI 4.8 形式已被纯化 95 倍，达到每毫克蛋白质 1755 纳摩尔/分钟的比活性。纯化程序包括离子交换色谱、凝胶过滤和亲和色谱。后者包括蓝琼脂糖亲和色谱和谷胱甘肽 - PGA1 - 琼脂糖亲和色谱。纯化的酶制剂还显示出较弱的依赖 NADP⁺的 15 - 羟基前列腺素脱氢酶活性，以 PGE1 为底物时为每毫克蛋白质 20 纳摩尔/分钟。该脱氢酶的 Km（PGE1）为 142.6 ± 45.1 微摩尔（标准误，n = 7）。