Skip to main content
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1974 Apr;53(4):1124–1131. doi: 10.1172/JCI107650

The Mechanism of Activation of Hormone-Sensitive Lipase in Human Adipose Tissue

John C Khoo 1, Alegria A Aquino 1, Daniel Steinberg 1
PMCID: PMC333098  PMID: 4360857

Abstract

A partially purified hormone-sensitive triglyceride lipase of human adipose tissue was found to be activated twofold by the addition of cyclic 3′,5′-AMP, ATP, and magnesium ions. Lipase activities against diolein and monoolein were not affected. Addition of protein kinase inhibitor at zero time completely inhibited activation, and this inhibition was prevented by prior addition of an excess of exogenous protein kinase (from rabbit skeletal muscle). Addition of protein kinase inhibitor during the activation step blocked the activation process without a time lag, suggesting that protein kinase operates directly on hormone-sensitive lipase. Further purification yielded a fraction free of protein kinase, and lipase activation in this fraction depended absolutely on addition of exogenous kinase. Incubation of human fat with epinephrine or isoproterenol stimulated lipolysis and caused conversion of nonactivated hormone-sensitive lipase to its activated form, as indicated by a decrease in the activation subsequently obtainable in fractions prepared from such hormone-treated tissues. These findings strongly suggest that the stimulation of lipolysis by hormonal treatment is the consequence of the activation of hormone-sensitive triglyceride lipase by cyclic 3′,5′-AMP-dependent protein kinase.

Full text

PDF
1128

Selected References

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

  1. Brown W. V., Baginsky M. L. Inhibition of lipoprotein lipase by an apoprotein of human very low density lipoprotein. Biochem Biophys Res Commun. 1972 Jan 31;46(2):375–382. doi: 10.1016/s0006-291x(72)80149-9. [DOI] [PubMed] [Google Scholar]
  2. Burns T. W., Langley P. E. Lipolysis by human adipose tissue: the role of cyclic 3',5'-adenosine monophosphate and adrenergic receptor sites. J Lab Clin Med. 1970 Jun;75(6):983–997. [PubMed] [Google Scholar]
  3. Burns T. W., Langley P. E., Robison G. A. Adrenergic receptors and cyclic AMP in the regulation of human adipose tissue lipolysis. Ann N Y Acad Sci. 1971 Dec 30;185:115–128. doi: 10.1111/j.1749-6632.1971.tb45242.x. [DOI] [PubMed] [Google Scholar]
  4. Butcher R. W., Ho R. J., Meng H. C., Sutherland E. W. Adenosine 3',5'-monophosphate in biological materials. II. The measurement of adenosine 3',5'-monophosphate in tissues and the role of the cyclic nucleotide in the lipolytic response of fat to epinephrine. J Biol Chem. 1965 Nov;240(11):4515–4523. [PubMed] [Google Scholar]
  5. Corbin J. D., Brostrom C. O., Alexander R. L., Krebs E. G. Adenosine 3',5'-monophosphate-dependent protein kinase from adipose tissue. J Biol Chem. 1972 Jun 25;247(12):3736–3743. [PubMed] [Google Scholar]
  6. Corbin J. D., Reimann E. M., Walsh D. A., Krebs E. G. Activation of adipose tissue lipase by skeletal muscle cyclic adenosine 3',5'- monophosphate-stimulated protein kinase. J Biol Chem. 1970 Sep 25;245(18):4849–4851. [PubMed] [Google Scholar]
  7. DOLE V. P. A relation between non-esterified fatty acids in plasma and the metabolism of glucose. J Clin Invest. 1956 Feb;35(2):150–154. doi: 10.1172/JCI103259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Galton D. J., Bray G. A. Studies on lipolysis in human adipose cells. J Clin Invest. 1967 Apr;46(4):621–629. doi: 10.1172/JCI105564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Heller R. A., Steinberg D. Partial glyceridase activity of a protein-kinase activatable triglyceride lipase from rat adipose tissue. Biochim Biophys Acta. 1972 May 23;270(1):65–73. doi: 10.1016/0005-2760(72)90178-6. [DOI] [PubMed] [Google Scholar]
  10. Huttunen J. K., Steinberg D. Activation and phosphorylation of purified adipose tissue hormone-sensitive lipase by cyclic AMP-dependent protein kinase. Biochim Biophys Acta. 1971 Sep 1;239(3):411–427. doi: 10.1016/0005-2760(71)90034-8. [DOI] [PubMed] [Google Scholar]
  11. Huttunen J. K., Steinberg D., Mayer S. E. ATP-dependent and cyclic AMP-dependent activation of rat adipose tissue lipase by protein kinase from rabbit skeletal muscle. Proc Natl Acad Sci U S A. 1970 Sep;67(1):290–295. doi: 10.1073/pnas.67.1.290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kelley T. F. Rapid assay of labeled free fatty acids in mixtures of labeled lipids. J Lipid Res. 1968 Nov;9(6):799–800. [PubMed] [Google Scholar]
  13. Khoo J. C., Fong W. W., Steinberg D. Activation of hormone-sensitive lipase from adipose tissue by cyclic AMP-dependent protein kinase. Biochem Biophys Res Commun. 1972 Oct 17;49(2):407–413. doi: 10.1016/0006-291x(72)90425-1. [DOI] [PubMed] [Google Scholar]
  14. Krebs E. G. Protein kinases. Curr Top Cell Regul. 1972;5:99–133. [PubMed] [Google Scholar]
  15. 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]
  16. Manganiello V. C., Murad F., Vaughan M. Effects of lipolytic and antilipolytic agents on cyclic 3',5'-adenosine monophosphate in fat cells. J Biol Chem. 1971 Apr 10;246(7):2195–2202. [PubMed] [Google Scholar]
  17. Mosinger B., Kuhn E., Kujalová V. Action of adipokinetic hormones on human adipose tissue in vitro. J Lab Clin Med. 1965 Sep;66(3):380–389. [PubMed] [Google Scholar]
  18. Moskowitz J., Fain J. N. Hormonal regulation of lipolysis and phosphorylase activity in human fat cells. J Clin Invest. 1969 Oct;48(10):1802–1808. doi: 10.1172/JCI106146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nikkilä E. A., Pykälistö O. Regulation of adipose tissue lipoprotein lipase synthesis by intracellular free fatty acid. Life Sci. 1968 Dec 15;7(24):1303–1309. doi: 10.1016/0024-3205(68)90260-9. [DOI] [PubMed] [Google Scholar]
  20. Patten R. L. The reciprocal regulation of lipoprotein lipase activity and hormone-sensitive lipase activity in rat adipocytes. J Biol Chem. 1970 Nov 10;245(21):5577–5584. [PubMed] [Google Scholar]
  21. RIZACK M. A. ACTIVATION OF AN EPINEPHRINE-SENSITIVE LIPOLYTIC ACTIVITY FROM ADIPOSE TISSUE BY ADENOSINE 3',5'-PHOSPHATE. J Biol Chem. 1964 Feb;239:392–395. [PubMed] [Google Scholar]
  22. Sidhu K. S., Emery R. S. Regulation of blood fatty acids and glycerol in lactating cows. J Dairy Sci. 1972 Jul;55(7):926–930. doi: 10.3168/jds.S0022-0302(72)85598-X. [DOI] [PubMed] [Google Scholar]
  23. VAUGHAN M., STEINBERG D. EFFECT OF HORMONES ON LIPOLYSIS AND ESTERIFICATION OF FREE FATTY ACIDS DURING INCUBATION OF ADIPOSE TISSUE IN VITRO. J Lipid Res. 1963 Apr;4:193–199. [PubMed] [Google Scholar]
  24. Walsh D. A., Ashby C. D., Gonzalez C., Calkins D., Fischer E. H. Krebs EG: Purification and characterization of a protein inhibitor of adenosine 3',5'-monophosphate-dependent protein kinases. J Biol Chem. 1971 Apr 10;246(7):1977–1985. [PubMed] [Google Scholar]
  25. Walsh D. A., Perkins J. P., Krebs E. G. An adenosine 3',5'-monophosphate-dependant protein kinase from rabbit skeletal muscle. J Biol Chem. 1968 Jul 10;243(13):3763–3765. [PubMed] [Google Scholar]
  26. Weiss B., Davies J. I., Brodie B. B. Evidence for a role of adenosine 3',5'-monophosphate in adipose tissue lipolysis. Biochem Pharmacol. 1966 Oct;15(10):1553–1561. doi: 10.1016/0006-2952(66)90199-7. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES