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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Format: | Article |
Language: | English |
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Elsevier
1981-01-01
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Series: | Journal of Lipid Research |
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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. |
first_indexed | 2024-12-17T07:25:21Z |
format | Article |
id | doaj.art-191a236712f54465a12b62b2857a4e84 |
institution | Directory Open Access Journal |
issn | 0022-2275 |
language | English |
last_indexed | 2024-12-17T07:25:21Z |
publishDate | 1981-01-01 |
publisher | Elsevier |
record_format | Article |
series | Journal of Lipid Research |
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 |