Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1975 Jun;147(3):531–539. doi: 10.1042/bj1470531

Selective inhibition of cholesterol synthesis by cell-free preparations of rat liver by using inhibitors of cytoplasmic acetoacetyl-coenzyme A thiolase.

D P Bloxham
PMCID: PMC1165481  PMID: 241330

Abstract

Cytoplasmic acetoacetyl-CoA thiolase (acetyl-CoA C-acetyltransferase, EC 2.3.1.9) was partially purified from rat liver. The enzyme was irreversibly inactivated by 4-bromocrotonyl-CoA, but-3-ynoyl-CoA, pent-3-ynoyl-CoA and dec-3-ynoyl-CoA. In the case of the alk-3-ynoyl-CoA esters the potency as alkylating agents of acetoacetyl-CoA thiolase decreased with increased chain length of the alk-3-ynoyl moiety. Advantage was taken of the specific action of alk-3-ynoyl-CoA esters on acetoacetyl-CoA thiolase to show that in a postmitochondrial fraction from rat liver they are effective inhibitors of cholesterol synthesis from sodium [2-14C]acetate under conditions when mevalonate conversion into cholesterol and fatty acid synthesis are unafffected. Short-chain alk-3-ynoic acids have little effect on sterol synthesis, although dec-3-ynoic acid is an effective inhibitor owing to its conversion into the CoA ester by the microsomal fatty acyl-CoA synthetase.

Full text

PDF
531

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abeles R. H., Walsh C. T. Acetylenic enzyme inactivators. Inactivation of gamma-cystathionase, in vitro and in vivo, by propargylglycine. J Am Chem Soc. 1973 Sep 5;95(18):6124–6125. doi: 10.1021/ja00799a053. [DOI] [PubMed] [Google Scholar]
  2. Akhtar M., Rahimtula A. D., Watkinson I. A., Wilton D. C., Munday K. A. The status of C-6, C-7, C-15 and C-16 hydrogen atoms in cholesterol biosynthesis. Eur J Biochem. 1969 May 1;9(1):107–111. doi: 10.1111/j.1432-1033.1969.tb00582.x. [DOI] [PubMed] [Google Scholar]
  3. BRODIE J. D., WASSON G., PORTER J. W. The participation of malonyl coenzyme A in the biosynthesis of mevalonic acid. J Biol Chem. 1963 Apr;238:1294–1301. [PubMed] [Google Scholar]
  4. BUCHER N. L., OVERATH P., LYNEN F. beta-Hydroxy-beta-methyl-glutaryl coenzyme A reductase, cleavage and condensing enzymes in relation to cholesterol formation in rat liver. Biochim Biophys Acta. 1960 Jun 3;40:491–501. doi: 10.1016/0006-3002(60)91390-1. [DOI] [PubMed] [Google Scholar]
  5. Bar-Tana J., Rose G., Shapiro B. The purification and properties of microsomal palmitoyl-coenzyme A synthetase. Biochem J. 1971 Apr;122(3):353–362. doi: 10.1042/bj1220353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Barth C. A., Hackenschmidt H. J., Weis E. E., Decker K. F. Influence of kynuremate on cholesterol and fatty acid synthesis in isolated perfused rat liver. J Biol Chem. 1973 Jan 25;248(2):738–739. [PubMed] [Google Scholar]
  7. Bloxham D. P., Akhtar M. Studies on the control of cholesterol biosynthesis: the adenosine 3':5'-cyclic monophosphate-dependent accumulation of a steroid carboxylic acid. Biochem J. 1971 Jun;123(2):275–278. doi: 10.1042/bj1230275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bloxham D. P., Wilton D. C., Akhtar M. Studies on the mechanism and regulation of C-4 demethylation in cholesterol biosynthesis. The role of adenosine 3':5'-cyclic monophosphate. Biochem J. 1971 Nov;125(2):625–634. doi: 10.1042/bj1250625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Bradshaw R. A., Robinson G. W., Hass G. M., Hill R. L. The reaction of fumarase with iodoacetate and 4-bromocrotonate. J Biol Chem. 1969 Apr 10;244(7):1755–1763. [PubMed] [Google Scholar]
  10. Clinkenbeard K. D., Sugiyama T., Moss J., Reed W. D., Lane M. D. Molecular and catalytic properties of cytosolic acetoacetyl coenzyme A thiolase from avian liver. J Biol Chem. 1973 Apr 10;248(7):2275–2284. [PubMed] [Google Scholar]
  11. Dempsey M. E. Regulation of steroid biosynthesis. Annu Rev Biochem. 1974;43(0):967–990. doi: 10.1146/annurev.bi.43.070174.004535. [DOI] [PubMed] [Google Scholar]
  12. ELLMAN G. L. Tissue sulfhydryl groups. Arch Biochem Biophys. 1959 May;82(1):70–77. doi: 10.1016/0003-9861(59)90090-6. [DOI] [PubMed] [Google Scholar]
  13. Endo K., Helmkamp G. M., Jr, Bloch K. Mode of inhibition of beta-hydroxydecanoyl thioester dehydrase by 3-decynoyl-N-acetylcysteamine. J Biol Chem. 1970 Sep 10;245(17):4293–4296. [PubMed] [Google Scholar]
  14. Fimognari G. M., Rodwell V. W. Mevalonate biosynthesis in rat liver. Lipids. 1970 Jan;5(1):104–108. doi: 10.1007/BF02531103. [DOI] [PubMed] [Google Scholar]
  15. Gehring U., Riepertinger C., Lynen F. Reinigung und Kristallisation der Thiolase, Untersuchungen zum Wirkungsmechanismus. Eur J Biochem. 1968 Nov;6(2):264–280. doi: 10.1111/j.1432-1033.1968.tb00446.x. [DOI] [PubMed] [Google Scholar]
  16. HERNDON J. H., Jr, SIPERSTEIN M. D. Desmosterol deposition in human and experimental atherosclerosis. Circ Res. 1963 Feb;12:228–234. doi: 10.1161/01.res.12.2.228. [DOI] [PubMed] [Google Scholar]
  17. Hevey R. C., Babson J., Maycock A. L., Abeles R. H. Highly specific enzyme inhibitors. Inhibition of plasma amine oxidase. J Am Chem Soc. 1973 Sep 5;95(18):6125–6127. doi: 10.1021/ja00799a054. [DOI] [PubMed] [Google Scholar]
  18. Higgins M. J., Kekwick R. G. An investigation into the role of malonyl-coenzyme A in isoprenoid biosynthesis. Biochem J. 1973 May;134(1):295–310. doi: 10.1042/bj1340295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Holland P. C., Clark M. G., Bloxham D. P. Inactivation of pig heart thiolase by 3-butynoyl coenzyme A, 3-pentynoyl coenzyme A, and 4-bromocrotonyl coenzyme A. Biochemistry. 1973 Aug 14;12(17):3309–3315. doi: 10.1021/bi00741a024. [DOI] [PubMed] [Google Scholar]
  20. Holland P. C., Senior A. E., Sherratt H. S. Biochemical effects of the hypoglycaemic compound pent-4-enoic acid and related non-hypoglycaemic fatty acids. Effects of their coenzyme A esters on enzymes of fatty acid oxidation. Biochem J. 1973 Sep;136(1):173–184. doi: 10.1042/bj1360173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Holland P. C., Sherratt H. S. Biochemical effects of the hypoglycaemic compound pent-4-enoic acid and related non-hypoglycaemic fatty acids. Effects of the free acids and their carnitine esters on coenzyme A-dependent oxidations in rat liver mitochondria. Biochem J. 1973 Sep;136(1):157–171. doi: 10.1042/bj1360157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. KITZ R., WILSON I. B. Esters of methanesulfonic acid as irreversible inhibitors of acetylcholinesterase. J Biol Chem. 1962 Oct;237:3245–3249. [PubMed] [Google Scholar]
  23. KORNBERG A., PRICER W. E., Jr Enzymatic synthesis of the coenzyme A derivatives of long chain fatty acids. J Biol Chem. 1953 Sep;204(1):329–343. [PubMed] [Google Scholar]
  24. LYNEN F. Functional group of coenzyme A and its metabolic relations, especially in the fatty acid cycle. Fed Proc. 1953 Sep;12(3):683–691. [PubMed] [Google Scholar]
  25. Middleton B. The kinetic mechanism and properties of the cytoplasmic acetoacetyl-coenzyme A thiolase from rat liver. Biochem J. 1974 Apr;139(1):109–121. doi: 10.1042/bj1390109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Middleton B. The oxoacyl-coenzyme A thiolases of animal tissues. Biochem J. 1973 Apr;132(4):717–730. doi: 10.1042/bj1320717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Middleton B., Tubbs P. K. An enzyme-bound intermediate in the biosynthesis of 3-hydroxy-3-methylglutaryl-coenzyme A. Biochem J. 1974 Jan;137(1):15–23. doi: 10.1042/bj1370015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Morisaki M., Bloch K. On the mode of interaction of -hydroxydecanoyl thioester dehydrase with allenic acid derivatives. Biochemistry. 1972 Feb 1;11(3):309–314. doi: 10.1021/bi00753a001. [DOI] [PubMed] [Google Scholar]
  29. Wheather D. W., Snow G. A. Assay of the mycobactins by measurement of the growth of Mycobacterium johnei. Biochem J. 1966 Jul;100(1):47–49. doi: 10.1042/bj1000047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Williamson J. R., Rostand S. G., Peterson M. J. Control factors affecting gluconeogenesis in perfused rat liver. Effects of 4-pentenoic acid. J Biol Chem. 1970 Jun;245(12):3242–3251. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES