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 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.
D000214:Acyl Coenzyme A / Q000302:isolation & purification*; D002848:Chromatography, DEAE-Cellulose; D005977:Glutarates / Q000302:isolation & purification*; D015093:Meglutol / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000302:isolation & purification*
D000214:Acyl Coenzyme A; D005977:Glutarates; C008047:3-hydroxy-3-methylglutaryl-coenzyme A; C029727:3-hydroxy-3-methylglutaric anhydride; D015093:Meglutol