Stable carbon isotope discrimination by human 3-hydroxy-3-methylglutaryl-coenzyme A reductase

被引：3

作者：

Luedke, Svena M. ^{[1
]}

Flenker, Ulrich ^{[1
]}

Schaenzer, Wilhelm ^{[1
]}

Schomburg, Dietmar ^{[2
]}

机构：

[1] German Sport Univ Cologne, Inst Biochem, D-50933 Cologne, Germany

[2] Tech Univ Carolo Wilhelmina Braunschweig, Inst Biochem & Biotechnol, D-38106 Braunschweig, Germany

来源：

JOURNAL OF LIPID RESEARCH | 2008年 / 49卷 / 12期

关键词：

mevalonate pathway; steroids; kinetic isotope effect; isotope ratio mass spectrometry; curve fitting; doping control;

D O I：

10.1194/jlr.M800313-JLR200

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The aim of this study was to investigate the possible existence and magnitude of stable carbon isotope discrimination by human 3-hydroxy-3-methylglutaryl-coenzyme A reductase ( HMGR). The catalytic portion of HMGR was expressed and purified. The reaction product mevalonate was lactonized and extracted from the reaction mixture by a solid-phase extraction protocol. Stable carbon isotope ratios of mevalonolactone ( MVL) were analyzed by gas chromatography-combustion-isotope ratio mass spectrometry. An average fractionation factor (12)k/(13)k of 1.0031 +/- 0.0004 for all carbon atoms contained in MVL was estimated by the method of internal competition. The value was calculated by nonlinear curve fitting, where the ratio C-13/C-12 of MVL was plotted versus the fraction of reaction. - Ludke, S. M., U. Flenker, W. Schanzer, and D. Schomburg. Stable carbon isotope discrimination by human 3-hydroxy-3-methylglutaryl-coenzyme A reductase. J. Lipid Res. 2008. 49: 2620-2626.

引用

页码：2620 / 2626

页数：7

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