Journal of Lipid Research | |
Stable carbon isotope discrimination by human 3-hydroxy-3-methylglutaryl-coenzyme A reductase | |
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