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Journal of Lipid Research, Vol 22, 184-187, Copyright © 1981 by Lipid Research, Inc.
ARTICLES |
IP Williamson and VW Rodwell
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 HMG0CoA.
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