Kinetic properties of hexose-monophosphate dehydrogenases. I. Isolation and partial purification of glucose-6-phosphate dehydrogenase from rat liver and kidney cortex.
Kinetic properties of hexose-monophosphate dehydrogenases. I. Isolation and partial purification of glucose-6-phosphate dehydrogenase from rat liver and kidney cortex.
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单磷酸己糖脱氢酶的动力学特性。
DOI：
10.1016/0024-3205(94)00433-s
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发表时间：
1995
期刊：
影响因子：
6.1
通讯作者：
J. A. Lupiáñez
中科院分区：
文献类型：
--
作者：
F. J. Corpas;L. García;J. Peragón;J. A. Lupiáñez
来自大鼠肝脏和肾皮质细胞质的葡萄糖 6-磷酸脱氢酶 (G6PDH) 已被部分纯化，并且几乎完全与 6-磷酸葡萄糖酸脱氢酶活性分离。纯化和分离程序包括高速离心、40-55% 硫酸铵分级分离，从而分离两种酶活性，最后将蛋白质级分应用到用 10 mM Tris-EDTA-NADP 缓冲液（pH 7.6）平衡的 Sephadex G-25 柱上，以消除任何污染代谢物。检查了离体肝脏和肾脏 G6PDH 的动力学特性。两种酶都显示出典型的米氏动力学饱和曲线，没有协同作用的证据。肝肾皮质G6PDH的最适pH均为9.4。 6-磷酸葡萄糖 (G6P) 和 NADP 的 Km 值分别为 3.29 × 10−4M 和 1.00 × 10−4M。在 37°C 和最适 pH 条件下测得的比活性为 327.1 mU/mg 蛋白质。 NADPH 用 Kiof 10 μM 引起竞争性抑制。肾皮质 G6PDH 的 G6P 和 NADP 的 Km 值分别为 2.06 × 10−4 和 0.25 × 10−4M。 pH 9.4 和 37°C 下的比活性为 76.55 mU/mg 蛋白质。 NADPH 抑制的 Ki 值为 4 μM。这项工作描述了一种简单、快速和可靠的方法，用于分离动物组织中参与己糖-单磷酸分流的两种脱氢酶。
Glucose-6-phosphate dehydrogenase (G6PDH) from rat-liver and kidney-cortex cytosol has been partially purified and almost completely separated from 6-phosphogluconate dehydrogenase activity. The purification and isolation procedures included high-speed centrifugation, 40–55% ammonium sulphate fractionation, by which both enzyme activities were separated, and finally, the application of the protein fraction to a column of Sephadex G-25 equilibrated with 10 mM Tris-EDTA-NADP buffer, pH 7.6, to eliminate any contaminating metabolites. The kinetic properties of isolated liver and renal G6PDH were examined. Both enzymes showed a typical Michaelis-Menten kinetic saturation curve with no evidence of co-operativity. The optimum pH of both liver and kidney cortex G6PDH was 9.4. The Km values for glucose-6-phosphate (G6P) and for NADP were 3.29 × 10−4M and 1.00 × 10−4M respectively. The specific activity measured at 37°C and optimum pH was 327.1 mU/ mg of protein. NADPH caused a competitive inhibition with a Kiof 10 μM. The Kmvalues for the G6P and NADP of kidney-cortex G6PDH were 2.06 × 10−4and 0.25 × 10−4M respectively. The specific activity at pH 9.4 and 37°C was 76.55 mU/ mg of protein. The Kivalue for NADPH inhibition was 4 μM. This work describes an easy, rapid and reliable method for the separation of the two dehydrogenases involved in the hexose-monophosphate shunt in animal tissues.
表型多样性是肝癌发生过程中假定的癌前肝细胞群的早期特性。
影响因子：
11.2
作者：
Ogawa,K;Solt,DB;Farber,E
通讯作者：
Farber,E
人红细胞中葡萄糖-6-磷酸脱氢酶的调节。
DOI：
--
发表时间：
1986
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Kirkman,HN;Gaetani,GF
通讯作者：
Gaetani,GF
氧化应激下牛睫状体己糖单磷酸分流的刺激。
DOI：
10.1089/jop.1986.2.59
发表时间：
1986
期刊：
Journal of ocular pharmacology
影响因子：
--
作者：
Shichi,H;Hodder,WA;Giblin,FJ
通讯作者：
Giblin,FJ