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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1981 Jan;78(1):120–123. doi: 10.1073/pnas.78.1.120

Specific phospholipids are required to reconstitute adenylate cyclase solubilized from rat brain.

G M Hebdon, H LeVine 3rd, N E Sahyoun, C J Schmitges, P Cuatrecasas
PMCID: PMC319002  PMID: 6941240

Abstract

Adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] was solubilized from a rat brain homogenate with sodium deoxycholate. This solubilized preparation had no detectable enzymic activity with either Mg-ATP or Mn-ATP as substrate. The activity could be restored by addition of either nonionic detergent or certain specific phospholipids. Maximal restoration of enzyme activity was obtained with Triton X-100, L-alpha-phosphatidylcholine, L-alpha-lysophosphatidylcholine, phosphatidyl-N-monomethylethanolamine, or sphingomyelin. Activity was only partially restored by phosphatidylethanolamine (40-60%) or phosphatidyl-N,N-dimethylethanolamine (10-20%). Other phospholipids tested, including phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, and phosphatidic acid, could not restore enzyme activity but, instead, could inhibit the stimulation of enzyme activity by phosphatidylcholine. The restoration of activity by L-alpha-phosphatidylcholine was inhibited by cholesterol at concentrations above 33 mol %, although this effect was not observed with three different esters of cholesterol. These studies suggest a possible specific role of phospholipids in modulating adenylate cyclase activity.

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Selected References

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

  1. Bakardjieva A., Galla H. J., Helmreich E. J. Modulation of the beta-receptor adenylate cyclase interactions in cultured Chang liver cells by phospholipid enrichment. Biochemistry. 1979 Jul 10;18(14):3016–3023. doi: 10.1021/bi00581a017. [DOI] [PubMed] [Google Scholar]
  2. Coleman R. Membrane-bound enzymes and membrane ultrastructure. Biochim Biophys Acta. 1973 Apr 3;300(1):1–30. doi: 10.1016/0304-4157(73)90010-5. [DOI] [PubMed] [Google Scholar]
  3. Drummond G. I., Duncan L. Adenyl cyclase in cardiac tissue. J Biol Chem. 1970 Mar 10;245(5):976–983. [PubMed] [Google Scholar]
  4. Engelhard V. H., Glaser M., Storm D. R. Effect of membrane phospholipid compositional changes on adenylate cyclase in LM cells. Biochemistry. 1978 Aug 8;17(16):3191–3200. doi: 10.1021/bi00609a004. [DOI] [PubMed] [Google Scholar]
  5. Fry M., Green D. E. Cardiolipin requirement by cytochrome oxidase and the catalytic role of phospholipid. Biochem Biophys Res Commun. 1980 Apr 29;93(4):1238–1246. doi: 10.1016/0006-291x(80)90622-1. [DOI] [PubMed] [Google Scholar]
  6. Hebdon G. M., Le Vine H., 3rd, Minard R. B., Sahyoun N. E., Schmitges C. J., Cuatrecasas P. Incorporation of rat brain adenylate cyclase into artificial phospholipid vesicles. J Biol Chem. 1979 Oct 25;254(20):10459–10465. [PubMed] [Google Scholar]
  7. Hirata F., Axelrod J. Enzymatic methylation of phosphatidylethanolamine increases erythrocyte membrane fluidity. Nature. 1978 Sep 21;275(5677):219–220. doi: 10.1038/275219a0. [DOI] [PubMed] [Google Scholar]
  8. Hirata F., Strittmatter W. J., Axelrod J. beta-Adrenergic receptor agonists increase phospholipid methylation, membrane fluidity, and beta-adrenergic receptor-adenylate cyclase coupling. Proc Natl Acad Sci U S A. 1979 Jan;76(1):368–372. doi: 10.1073/pnas.76.1.368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Houslay M. D., Warren G. B., Birdsall N. J., Metcalfe J. C. Lipid phase transitions control beta-hydroxybutyrate dehydrogenase activity in defined-lipid protein complexes. FEBS Lett. 1975 Mar 1;51(1):146–151. doi: 10.1016/0014-5793(75)80873-8. [DOI] [PubMed] [Google Scholar]
  10. Klein I., Moore L., Pastan I. Effect of liposomes containing cholesterol on adenylate cyclase activity of cultured mammalian fibroblasts. Biochim Biophys Acta. 1978 Jan 4;506(1):42–53. doi: 10.1016/0005-2736(78)90433-9. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Londos C., Lad P. M., Nielsen T. B., Rodbell M. Solubilization and conversion of hepatic adenylate cyclase to a form requiring MnATP as substrate. J Supramol Struct. 1979;10(1):31–37. doi: 10.1002/jss.400100104. [DOI] [PubMed] [Google Scholar]
  13. Robison G. A., Butcher R. W., Sutherland E. W. Cyclic AMP. Annu Rev Biochem. 1968;37:149–174. doi: 10.1146/annurev.bi.37.070168.001053. [DOI] [PubMed] [Google Scholar]
  14. SEKUZU I., JURTSHUK P., Jr, GREEN D. E. Studies on the electron transfer system. LI. Isolation and characterization of the D-(--)-beta-hydroxybutyric apodehydrogenase from beef heart mitochondria. J Biol Chem. 1963 Mar;238:975–982. [PubMed] [Google Scholar]
  15. SUTHERLAND E. W., RALL T. W., MENON T. Adenyl cylase. I. Distribution, preparation, and properties. J Biol Chem. 1962 Apr;237:1220–1227. [PubMed] [Google Scholar]
  16. Sandermann H., Jr Regulation of membrane enzymes by lipids. Biochim Biophys Acta. 1978 Sep 29;515(3):209–237. doi: 10.1016/0304-4157(78)90015-1. [DOI] [PubMed] [Google Scholar]
  17. Sinensky M., Minneman K. P., Molinoff P. B. Increased membrane acyl chain ordering activates adenylate cyclase. J Biol Chem. 1979 Sep 25;254(18):9135–9141. [PubMed] [Google Scholar]
  18. Tornqvist H., Belfrage P. Determination of protein in adipose tissue extracts. J Lipid Res. 1976 Sep;17(5):542–545. [PubMed] [Google Scholar]
  19. Warren G. B., Toon P. A., Birdsall N. J., Lee A. G., Metcalfe J. C. Reversible lipid titrations of the activity of pure adenosine triphosphatase-lipid complexes. Biochemistry. 1974 Dec 31;13(27):5501–5507. doi: 10.1021/bi00724a008. [DOI] [PubMed] [Google Scholar]

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