Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1982 Apr 15;204(1):345–352. doi: 10.1042/bj2040345

Reversibility of the insulin-stimulated phosphorylation of ATP citrate lyase and a cytoplasmic protein of subunit Mr 22000 in adipose tissue.

G J Belsham, R W Brownsey, R M Denton
PMCID: PMC1158351  PMID: 7052067

Abstract

Exposure of rat epididymal adipose tissue to insulin leads to a 2-fold increase in incorporation of [32P]phosphate into ATP citrate lyase and a cytoplasmic protein of Mr 22000. A smaller increase and decrease in incorporation into cytoplasmic proteins of Mr 63000 and 20000 respectively were also observed. Subsequent addition of anti-insulin serum largely reversed these changes within 15 min, indicating that the alterations in 32P incorporation represented changes in net phosphate content of the proteins.

Full text

PDF

Images in this article

348Fig. 1.
on p.348

Selected References

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

  1. Alexander M. C., Kowaloff E. M., Witters L. A., Dennihy D. T., Avruch J. Purification of a hepatic 123,000-dalton hormone-stimulated 32P-peptide and its identification as ATP-citrate lyase. J Biol Chem. 1979 Aug 25;254(16):8052–8056. [PubMed] [Google Scholar]
  2. Avruch J., Leone G. R., Martin D. B. Effects of epinephrine and insulin on phosphopeptide metabolism in adipocytes. J Biol Chem. 1976 Mar 10;251(5):1511–1515. [PubMed] [Google Scholar]
  3. Avruch J., Leone G. R., Martin D. B. Identification and subcellular distribution of adipocyte peptides and phosphopeptides. J Biol Chem. 1976 Mar 10;251(5):1505–1510. [PubMed] [Google Scholar]
  4. Belsham G. J., Brownsey R. W., Hughes W. A., Denton R. M. Anti-insulin receptor antibodies mimic the effects of insulin on the activities of pyruvate dehydrogenase and acetylCoA carboxylase and on specific protein phosphorylation in rat epididymal fat cells. Diabetologia. 1980 Apr;18(4):307–312. doi: 10.1007/BF00251011. [DOI] [PubMed] [Google Scholar]
  5. Belsham G. J., Denton R. M., Tanner M. J. Use of a novel rapid preparation of fat-cell plasma membranes employing Percoll to investigate the effects of insulin and adrenaline on membrane protein phosphorylation within intact fat-cells. Biochem J. 1980 Nov 15;192(2):457–467. doi: 10.1042/bj1920457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Belsham G. J., Denton R. M. The effect of insulin and adrenaline on the phosphorylation of a 22 000-molecular weight protein within isolated fat cells; possible identification as the inhibitor-1 of the 'general phosphatase' [proceedings]. Biochem Soc Trans. 1980 Jun;8(3):382–383. doi: 10.1042/bst0080382. [DOI] [PubMed] [Google Scholar]
  7. Benjamin W. B., Clayton N. L. Action of insulin and catecholamines on the phosphorylation of proteins associated with the cytosol, membranes, and "fat cake" of rat fat cells. J Biol Chem. 1978 Mar 10;253(5):1700–1709. [PubMed] [Google Scholar]
  8. Benjamin W. B., Singer I. Actions of insulin, epinephrine, and dibutyryl cyclic adenosine 5'-monophosphate on fat cell protein phosphorylations. Cyclic adenosine 5'-monophosphate dependent and independent mechanisms. Biochemistry. 1975 Jul 29;14(15):3301–3309. doi: 10.1021/bi00686a003. [DOI] [PubMed] [Google Scholar]
  9. Benjamin W. B., Singer I. Effect of insulin on the phosphorylation of adipose tissue protein. Biochim Biophys Acta. 1974 May 10;351(1):28–41. doi: 10.1016/0005-2795(74)90063-4. [DOI] [PubMed] [Google Scholar]
  10. Brownsey R. W., Belsham G. J., Denton R. M. Evidence for phosphorylation and activation of acetyl CoA carboxylase by a membrane-associated cyclic AMP-independent protein kinase. Relationship to the activation of acetyl CoA carboxylase by insulin. FEBS Lett. 1981 Feb 23;124(2):145–150. doi: 10.1016/0014-5793(81)80123-8. [DOI] [PubMed] [Google Scholar]
  11. Brownsey R. W., Denton R. M. Evidence that insulin activates fat-cell acetyl-CoA carboxylase by increased phosphorylation at a specific site. Biochem J. 1982 Jan 15;202(1):77–86. doi: 10.1042/bj2020077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Brownsey R. W., Hughes W. A., Denton R. M. Adrenaline and the regulation of acetyl-coenzyme A carboxylase in rat epididymal adipose tissue. Inactivation of the enzyme is associated with phosphorylation and can be reversed on dephosphorylation. Biochem J. 1979 Oct 15;184(1):23–32. doi: 10.1042/bj1840023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Crapo L., Graham W., Avruch J. Studies on the mechanism of insulin-stimulated protein phosphorylation in adipocytes. Biochem Biophys Res Commun. 1980 Jun 30;94(4):1331–1336. doi: 10.1016/0006-291x(80)90565-3. [DOI] [PubMed] [Google Scholar]
  14. Denton R. M., Brownsey R. W., Belsham G. J. A partial view of the mechanism of insulin action. Diabetologia. 1981 Oct;21(4):347–362. doi: 10.1007/BF00252681. [DOI] [PubMed] [Google Scholar]
  15. Forn J., Greengard P. Regulation by lipolytic and antillipolytic compounds of the phosphorylation of specific proteins in isolated intact fat cells. Arch Biochem Biophys. 1976 Oct;176(2):721–733. doi: 10.1016/0003-9861(76)90216-2. [DOI] [PubMed] [Google Scholar]
  16. Foulkes J. G., Cohen P. The hormonal control of glycogen metabolism. Phosphorylation of protein phosphatase inhibitor-1 in vivo in response to adrenaline. Eur J Biochem. 1979 Jun;97(1):251–256. doi: 10.1111/j.1432-1033.1979.tb13109.x. [DOI] [PubMed] [Google Scholar]
  17. Huang F. L., Glinsmann W. H. Separation and characterization of two phosphorylase phosphatase inhibitors from rabbit skeletal muscle. Eur J Biochem. 1976 Nov 15;70(2):419–426. doi: 10.1111/j.1432-1033.1976.tb11032.x. [DOI] [PubMed] [Google Scholar]
  18. Hughes W. A., Brownsey R. W., Denton R. M. Studies on the incorporation of [32P]phosphate into pyruvate dehydrogenase in intact rat fat-cells. Effects of insulin. Biochem J. 1980 Nov 15;192(2):469–481. doi: 10.1042/bj1920469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Janski A. M., Srere P. A., Cornell N. W., Veech R. L. Phosphorylation of ATP citrate lyase in response to glucagon. J Biol Chem. 1979 Oct 10;254(19):9365–9368. [PubMed] [Google Scholar]
  20. Kahn C. R., Baird K. L., Jarrett D. B., Flier J. S. Direct demonstration that receptor crosslinking or aggregation is important in insulin action. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4209–4213. doi: 10.1073/pnas.75.9.4209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Karnieli E., Zarnowski M. J., Hissin P. J., Simpson I. A., Salans L. B., Cushman S. W. Insulin-stimulated translocation of glucose transport systems in the isolated rat adipose cell. Time course, reversal, insulin concentration dependency, and relationship to glucose transport activity. J Biol Chem. 1981 May 25;256(10):4772–4777. [PubMed] [Google Scholar]
  22. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  23. Lapetina E. G., Michell R. H. Phosphatidylinositol metabolism in cells receiving extracellular stimulation. FEBS Lett. 1973 Apr 1;31(1):1–10. doi: 10.1016/0014-5793(73)80061-4. [DOI] [PubMed] [Google Scholar]
  24. Laskey R. A. Molecular mechanisms of chromatin assembly. Biochem Soc Trans. 1981 Aug;9(4):263–270. doi: 10.1042/bst0090263. [DOI] [PubMed] [Google Scholar]
  25. Lastick S. M., McConkey E. H. HeLa ribosomal protein S6. Insulin and dibutyryl cyclic AMP affect different phosphopeptides. J Biol Chem. 1981 Jan 25;256(2):583–585. [PubMed] [Google Scholar]
  26. Lee K. H., Kim K. H. Stimulation by epinephrine of in vivo phosphorylation and inactivation of acetyl coenzyme A carboxylase of rat epididymal adipose tissue. J Biol Chem. 1979 Mar 10;254(5):1450–1453. [PubMed] [Google Scholar]
  27. McCormack J. G., Denton R. M. Evidence that fatty acid synthesis in the interscapular brown adipose tissue of cold-adapted rats is increased in vivo by insulin by mechanisms involving parallel activation of pyruvate dehydrogenase and acetyl-coenzyme A carboxylase. Biochem J. 1977 Sep 15;166(3):627–630. doi: 10.1042/bj1660627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Michell R. H. Inositol phospholipids and cell surface receptor function. Biochim Biophys Acta. 1975 Mar 25;415(1):81–47. doi: 10.1016/0304-4157(75)90017-9. [DOI] [PubMed] [Google Scholar]
  29. Ramakrishna S., Benjamin W. B. Phosphorylation of ATP-citrate lyase by a cyclic AMP-independent protein kinase from rat liver. FEBS Lett. 1981 Feb 23;124(2):140–144. doi: 10.1016/0014-5793(81)80122-6. [DOI] [PubMed] [Google Scholar]
  30. Smith C. J., Wejksnora P. J., Warner J. R., Rubin C. S., Rosen O. M. Insulin-stimulated protein phosphorylation in 3T3-L1 preadipocytes. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2725–2729. doi: 10.1073/pnas.76.6.2725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Stansbie D., Brownsey R. W., Crettaz M., Denton R. M. Acute effects in vivo of anti-insulin serum on rates of fatty acid synthesis and activities of acetyl-coenzyme A carboxylase and pyruvate dehydrogenase in liver and epididymal adipose tissue of fed rats. Biochem J. 1976 Nov 15;160(2):413–416. doi: 10.1042/bj1600413. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES