PURIFICATION OF A DISTINCTIVE FORM OF ENDOTOXIN-INDUCED NITRIC-OXIDE SYNTHASE FROM RAT-LIVER

被引：95

作者：

EVANS, T ^{[1
]}

CARPENTER, A ^{[1
]}

COHEN, J ^{[1
]}

机构：

[1] ROYAL POSTGRAD MED SCH,DEPT INFECT DIS,DU CANE RD,LONDON W12 ONN,ENGLAND

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 1992年 / 89卷 / 12期

关键词：

ENDOTHELIUM-DERIVED RELAXING FACTOR; L-ARGININE; CALMODULIN;

D O I：

10.1073/pnas.89.12.5361

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

An endotoxin-induced form of nitric oxide synthase (EC 1.14.23) was purified to homogeneity from rat liver by sequential anion-exchange chromatography and affinity chromatography using 2',5'-ADP-Sepharose. The enzyme has a subunit molecular mass of 135 kDa as determined by SDS/PAGE, a maximum specific activity of 462 nmol of citrulline formed from arginine per min per mg, and a K(m) for arginine of 11-mu-M. The enzyme was strongly stimulated by the addition of calmodulin with an EC50 of 2 nM, but removal of free calcium from the assay medium only reduced activity by 15%. Calmodulin inhibitors significantly reduced the enzyme activity. Tetrahydrobiopterin, FAD, and FMN were all required for full enzyme activity. This form of endotoxin-induced nitric oxide synthase from liver differs from the inducible enzyme found in macrophages and is unusual in that it is stimulated by calmodulin with little dependence on the calcium ion concentration.

引用

页码：5361 / 5365

页数：5

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