J. Lipid Res.
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Journal of Lipid Research, Vol 20, 40-46, Copyright © 1979 by Lipid Research, Inc.

ARTICLES

Improved methods for the solubilization and assay of hepatic 3-hydroxy- 3-methylglutaryl coenzyme A reductase

PA Edwards, D Lemongello and AM Fogelman

A method for solubilizing HMG-CoA reductase is described that reproducibly yielded approximately 190% of the activity assayed in rat liver microsomes. Optimal solubilization occurred when microsomal membranes were frozen at a fixed concentration, thawed, homogenized in a buffer containing 50% glycerol, and incubated at 37 degrees C for 60 minutes. A rapid spectrophotometric assay of the reductase has been developed and the optimal conditions defined. Using this assay, the kinetics were determined for HMG-CoA reductase purified to a specific activity of 17,400 nmol NADPH oxidized per minute per mg protein.
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