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

Journal of Lipid Research

Dietmar Schomburg¹  Svena M. Lüdke²  Ulrich Flenker²  Wilhelm Schänzer² 

[1] Institute for Biochemistry and Biotechnology, Technical University Braunschweig, Langer Kamp 19B, 38106 Braunschweig, Germany;Institute of Biochemistry, German Sport University Cologne, Am Sportpark Müngersdorf 6, 50933 Cologne, Germany;

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

DOI  :  

来源: DOAJ

【 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 12k/13k 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 13C/12C of MVL was plotted versus the fraction of reaction.

【 授权许可】

Unknown