Isolation and purification of 3-hydroxy-3-methylglutaryl-coenzyme A by ion-exchange chromatography

For precise determination of the catalytic activity of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase (EC 1.1.1.34), the HMG-CoA employed as substrate must be free of HMG, CoA, and other inhibitors of HMG-CoA reductase activity. The standard purification of HMG-CoA by paper chromatography...

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Main Authors: I P Williamson, V W Rodwell
Format: Article
Language:English
Published: Elsevier 1981-01-01
Series:Journal of Lipid Research
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S0022227520347556
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author I P Williamson
V W Rodwell
author_facet I P Williamson
V W Rodwell
author_sort I P Williamson
collection DOAJ
description For precise determination of the catalytic activity of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase (EC 1.1.1.34), the HMG-CoA employed as substrate must be free of HMG, CoA, and other inhibitors of HMG-CoA reductase activity. The standard purification of HMG-CoA by paper chromatography gives poor resolution of HMG-CoA from CoA and may be accompanied by some decomposition of HMG-CoA. We describe a simplified procedure for synthesis and for isolation from the reaction mixture of homogeneous, high specific activity [3(-14)C]HMG-CoA free of HMG, CoA, or nonpolar contaminants. Isolation of HMG-CoA utilizes ion-exchange chromatography in a gradient of ammonium formate, which is subsequently removed by lyophilization. The methods are proposed for use in the preparation or isolation of HMG-CoA.
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spelling doaj.art-191a236712f54465a12b62b2857a4e842022-12-21T21:58:39ZengElsevierJournal of Lipid Research0022-22751981-01-01221184187Isolation and purification of 3-hydroxy-3-methylglutaryl-coenzyme A by ion-exchange chromatographyI P Williamson0V W Rodwell1Depurtment of Biochemistry, Marischal College, Univrrsity of Aberdeen, Aberdeen, AB9 1AS, Scotland and Department qf Biochemistry, Purdue University, West Lafayette, IN 47907Department of Biochemistry, Purdue University, West Lafayette, IN 47907For precise determination of the catalytic activity of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase (EC 1.1.1.34), the HMG-CoA employed as substrate must be free of HMG, CoA, and other inhibitors of HMG-CoA reductase activity. The standard purification of HMG-CoA by paper chromatography gives poor resolution of HMG-CoA from CoA and may be accompanied by some decomposition of HMG-CoA. We describe a simplified procedure for synthesis and for isolation from the reaction mixture of homogeneous, high specific activity [3(-14)C]HMG-CoA free of HMG, CoA, or nonpolar contaminants. Isolation of HMG-CoA utilizes ion-exchange chromatography in a gradient of ammonium formate, which is subsequently removed by lyophilization. The methods are proposed for use in the preparation or isolation of HMG-CoA.http://www.sciencedirect.com/science/article/pii/S0022227520347556HMG-CoA reductasecholesterogenesismevalonolactone
spellingShingle I P Williamson
V W Rodwell
Isolation and purification of 3-hydroxy-3-methylglutaryl-coenzyme A by ion-exchange chromatography
Journal of Lipid Research
HMG-CoA reductase
cholesterogenesis
mevalonolactone
title Isolation and purification of 3-hydroxy-3-methylglutaryl-coenzyme A by ion-exchange chromatography
title_full Isolation and purification of 3-hydroxy-3-methylglutaryl-coenzyme A by ion-exchange chromatography
title_fullStr Isolation and purification of 3-hydroxy-3-methylglutaryl-coenzyme A by ion-exchange chromatography
title_full_unstemmed Isolation and purification of 3-hydroxy-3-methylglutaryl-coenzyme A by ion-exchange chromatography
title_short Isolation and purification of 3-hydroxy-3-methylglutaryl-coenzyme A by ion-exchange chromatography
title_sort isolation and purification of 3 hydroxy 3 methylglutaryl coenzyme a by ion exchange chromatography
topic HMG-CoA reductase
cholesterogenesis
mevalonolactone
url http://www.sciencedirect.com/science/article/pii/S0022227520347556
work_keys_str_mv AT ipwilliamson isolationandpurificationof3hydroxy3methylglutarylcoenzymeabyionexchangechromatography
AT vwrodwell isolationandpurificationof3hydroxy3methylglutarylcoenzymeabyionexchangechromatography