Purification of 3-hydroxy-3-methylglutaryl coenzyme A reductase.

The enzyme 3-hydroxy-3-methylglutaryl coenzyme agarose, glucose-6-phosphate dehydrogenase, and aldolA reductase from human liver was purified to homogeneity. ase were from Sigma Chemical co.; agarose-hexaneHMG-CoA was from P-L Biochemicals; bovine serum Electrophoresis of the purified human enzyme on non-denaturing gels revealed a single protein-staining band that co-migrated with reductase activity and could be selectively removed a1bumin was from Pentex; DL-3-hydroxy-3-methy1 i3by treatment with cross-reacting antibody prepared to the pu''C]glutaryl CoA and [5-3Hlmevalonic acid were from rified reductase from rat liver. On SDS polyacrylamide gel New England Nuclear; Dowex AGl-X8 formate, soelectrophoresis, the purified reductase showed only one proteinimately 52,000. Based on the sedimentation rate during sucrose density gradient centrifugation, most of the enzyme from crude solubilized preparations had an apparent molecular weight of approximately 104,000. The human reductase, like the rat liver enzyme, was dimeric and composed of subunits of approximately 50,000 molecular weight.-Tanaka R. D., P. A. Edwards, S. H. Lan, E. M. Kniippel, and A. M. Fogelman. Purification of 3-hydroxy-3-methylglutaryl coenzyme A reductase from human liver. J. Lipid Res. 1982. 2 3 523-530. Supplementary key words cholesterogenesis immunotitration dium dodecyl sulfate, and acrylamide were from Biohave been previously reported (5).