Abstract
A procedure for the purification of 3-hydroxy-3-methylglutaryl-coenzyme A reductase [mevalonate: NADP+ oxidoreductase (CoA-acylating); EC 1.1.1.34] solubilized from rat liver microsomes is reported. This enzyme has a specific activity of 9,000-10,000 nmol of mevalonate formed per min/mg of protein. This represents a 4100-fold purification over the activity in microsomes, and a specific activity that is approximately 20-fold greater than the highest previously reported value. The enzyme is judged to be homogeneous on the basis of sodium dodecyl sulfate/polyacrylamide disc gel electrophoresis, polyacrylamide disc gel electrophoresis, and immunoanalysis. Data are also presented that indicate the separation of enzymatically active and inactive species of 3-hydroxy-3-methylglutaryl-coenzyme A reductase on affinity chromatography on a coenzyme A column.
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ackerman M. E., Redd W. L., Scallen T. J. Solubilization of 3-hydroxy-3-methylglutaryl coenzyme A reductase from lyophilized rat liver microsomes: lack of evidence for cold lability in this soluble enzyme preparation. Biochem Biophys Res Commun. 1974 Jan;56(1):29–35. doi: 10.1016/s0006-291x(74)80311-6. [DOI] [PubMed] [Google Scholar]
- Beg Z. H., Allmann D. W., Gibson D. M. Modulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity with cAMP and wth protein fractions of rat liver cytosol. Biochem Biophys Res Commun. 1973 Oct 15;54(4):1362–1369. doi: 10.1016/0006-291x(73)91137-6. [DOI] [PubMed] [Google Scholar]
- Berndt J., Gaumert R. Evidence for an activating-inactivating system of 3-hydroxy-3-methylglutaryl-CoA reductase in mouse liver. Hoppe Seylers Z Physiol Chem. 1974 Jul;355(7):905–910. [PubMed] [Google Scholar]
- Brown M. S., Brunschede G. Y., Goldstein J. L. Inactivation of 3-hydroxy-3-methylglutaryl coenzyme A reductase in vitro. An adenine nucleotide-dependent reaction catalyzed by a factor in human fibroblasts. J Biol Chem. 1975 Apr 10;250(7):2502–2509. [PubMed] [Google Scholar]
- Brown M. S., Dana S. E., Dietschy J. M., Siperstein M. D. 3-Hydroxy-3-methylglutaryl coenzyme A reductase. Solubilization and purification of a cold-sensitive microsomal enzyme. J Biol Chem. 1973 Jul 10;248(13):4731–4738. [PubMed] [Google Scholar]
- Dunker A. K., Rueckert R. R. Observations on molecular weight determinations on polyacrylamide gel. J Biol Chem. 1969 Sep 25;244(18):5074–5080. [PubMed] [Google Scholar]
- Heller R. A., Gould R. G. Reversible cold inactivation of microsomal 3-hydroxy-3-methylglutaryl coenzyme A reductase from rat liver. J Biol Chem. 1974 Aug 25;249(16):5254–5260. [PubMed] [Google Scholar]
- Heller R. A., Gould R. G. Solubilization and partial purification of hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase. Biochem Biophys Res Commun. 1973 Feb 5;50(3):859–865. doi: 10.1016/0006-291x(73)91324-7. [DOI] [PubMed] [Google Scholar]
- Heller R. A., Shrewsbury M. A. 3-Hydroxy-3-methylglutaryl coenzyme A reductase from rat liver. Its purification, properties, and immunochemical studies. J Biol Chem. 1976 Jun 25;251(12):3815–3822. [PubMed] [Google Scholar]
- Higgins M. J., Brady D., Rudney H. Rat liver 3-hydroxy-3-methylglutaryl coenzyme A reductase: a comparison and immunological study of purified solubilized preparations, and alteration of enzyme levels by cholestyramine feeding. Arch Biochem Biophys. 1974 Jul;163(1):271–282. doi: 10.1016/0003-9861(74)90477-9. [DOI] [PubMed] [Google Scholar]
- Kawachi T., Rudney H. Solubilization and purification of beta-hydroxy-beta-methylglutaryl coenzyme A reductase from rat liver. Biochemistry. 1970 Apr 14;9(8):1700–1705. doi: 10.1021/bi00810a008. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Linn T. C. The effect of cholesterol feeding and fasting upon beta-hydroxy-beta-methylglutaryl coenzyme A reductase. J Biol Chem. 1967 Mar 10;242(5):990–993. [PubMed] [Google Scholar]
- Nepokroeff C. M., Lakshmanan M. R., Ness G. C., Dugan R. E., Porter J. W. Regulation of the diurnal rhythm of rat liver beta-hydroxy-beta-methylglutaryl coenzmye A reductase activity by insulin, glucagon, cyclic AMP and hydrocortisone. Arch Biochem Biophys. 1974 Feb;160(2):387–396. doi: 10.1016/0003-9861(74)90412-3. [DOI] [PubMed] [Google Scholar]
- Qureshi N., Dugan R. E., Nimmannit S., Wu W. H., Porter J. W. Purification of beta-hydroxy-beta-methylglutaryl-coenzyme A reductase from yeast. Biochemistry. 1976 Sep 21;15(19):4185–4190. doi: 10.1021/bi00664a009. [DOI] [PubMed] [Google Scholar]
- Shapiro D. J., Nordstrom J. L., Mitschelen J. J., Rodwell V. W., Schimke R. T. Micro assay for 3-hydroxy-3-methylglutaryl-CoA reductase in rat liver and in L-cell fibroblasts. Biochim Biophys Acta. 1974 Dec 29;370(2):369–377. doi: 10.1016/0005-2744(74)90098-9. [DOI] [PubMed] [Google Scholar]
- Shapiro D. J., Rodwell V. W. Fine structure of the cyclic rhythm of 3-hydroxy-3-methylglutaryl coenzyme A reductase. Differential effects of cholesterol feeding and fasting. Biochemistry. 1972 Mar 14;11(6):1042–1045. doi: 10.1021/bi00756a014. [DOI] [PubMed] [Google Scholar]
- Siperstein M. D., Fagan V. M. Feedback control of mevalonate synthesis by dietary cholesterol. J Biol Chem. 1966 Feb 10;241(3):602–609. [PubMed] [Google Scholar]
- Tormanen C. D., Redd W. L., Srikantaiah M. V., Scallen T. J. Purification of 3-hydroxy-3-methylglutaryl coenzyme A reductase. Biochem Biophys Res Commun. 1976 Feb 9;68(3):754–762. doi: 10.1016/0006-291x(76)91209-2. [DOI] [PubMed] [Google Scholar]
- Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
- White L. W., Rudney H. Regulation of 3-hydroxy-3-methylglutarate and mevalonate biosynthesis by rat liver homogenates. Effects of fasting, cholesterol feeding, and triton administration. Biochemistry. 1970 Jun 23;9(13):2725–2731. doi: 10.1021/bi00815a021. [DOI] [PubMed] [Google Scholar]




