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. 1981 Apr;78(4):2359–2362. doi: 10.1073/pnas.78.4.2359

Specific binding of solubilized adenylate cyclase to the erythrocyte cytoskeleton.

N E Sahyoun, H Le Vine 3rd, G M Hebdon, R Hemadah, P Cuatrecasas
PMCID: PMC319345  PMID: 6941294

Abstract

Concepts and criteria that have been developed for the study of the molecular organization of membrane-associated proteins are employed here to investigate the interaction of adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] with other membrane components. Detergent-solubilized adenylate cyclase can be shown to bind to erythrocyte-derived Triton X-100 shells containing cytoskeletal elements. This binding appears to be saturable with respect to adenylate cyclase concentration, and it is enhanced by the presence of divalent cations. Preactivation of the enzyme with 5'-guanylyl imidodiphosphate and isoproterenol, or with NaF, is a prerequisite for effective binding. Two exceptions to this general observation are noted: rat brain adenylate cyclase, which binds without prestimulation, and rat testicular cytosolic adenylate cyclase, which fails to bind under any of the conditions tried. The binding sites of the Triton X-100 shells are inactivated or released by treatment with various concentrations of trypsin or KCl. Moreover, exposure of the Triton X-100 shells to increasing temperatures results in a progressive loss of the adenylate cyclase binding capacity. On the basis of these and other findings, it is suggested that the adenylate cyclase complex possesses two principal domains that allow it to interact with both cytoskeletal elements and the lipid bilayer. The specific modulation of these interactions may be involved in the hormonal regulation of 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. Ash J. F., Louvard D., Singer S. J. Antibody-induced linkages of plasma membrane proteins to intracellular actomyosin-containing filaments in cultured fibroblasts. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5584–5588. doi: 10.1073/pnas.74.12.5584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bennett V., Branton D. Selective association of spectrin with the cytoplasmic surface of human erythrocyte plasma membranes. Quantitative determination with purified (32P)spectrin. J Biol Chem. 1977 Apr 25;252(8):2753–2763. [PubMed] [Google Scholar]
  3. Bennett V. Purification of an active proteolytic fragment of the membrane attachment site for human erythrocyte spectrin. J Biol Chem. 1978 Apr 10;253(7):2292–2299. [PubMed] [Google Scholar]
  4. Bennett V., Stenbuck P. J. Identification and partial purification of ankyrin, the high affinity membrane attachment site for human erythrocyte spectrin. J Biol Chem. 1979 Apr 10;254(7):2533–2541. [PubMed] [Google Scholar]
  5. Bennett V., Stenbuck P. J. The membrane attachment protein for spectrin is associated with band 3 in human erythrocyte membranes. Nature. 1979 Aug 9;280(5722):468–473. doi: 10.1038/280468a0. [DOI] [PubMed] [Google Scholar]
  6. Bergman R. N., Hechter O. Neurohypophyseal hormone-responsive renal adenylate cyclase. IV. A random-hit matrix model for coupline in a hormone-sensitive adenylate cyclase system. J Biol Chem. 1978 May 10;253(9):3238–3250. [PubMed] [Google Scholar]
  7. Blomberg F., Cohen R. S., Siekevitz P. The structure of postsynaptic densities isolated from dog cerebral cortex. II. Characterization and arrangement of some of the major proteins within the structure. J Cell Biol. 1977 Jul;74(1):204–225. doi: 10.1083/jcb.74.1.204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bourguignon L. Y., Singer S. J. Transmembrane interactions and the mechanism of capping of surface receptors by their specific ligands. Proc Natl Acad Sci U S A. 1977 Nov;74(11):5031–5035. doi: 10.1073/pnas.74.11.5031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Braun T., Dods R. F. Development of a Mn-2+-sensitive, "soluble" adenylate cyclase in rat testis. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1097–1101. doi: 10.1073/pnas.72.3.1097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cohen C. M., Branton D. The role of spectrin in erythrocyte membrane-stimulated actin polymerisation. Nature. 1979 May 10;279(5709):163–165. doi: 10.1038/279163a0. [DOI] [PubMed] [Google Scholar]
  11. Craig S. W., Cuatrecasas P. Mobility of cholera toxin receptors on rat lymphocyte membranes. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3844–3848. doi: 10.1073/pnas.72.10.3844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Flanagan J., Koch G. L. Cross-linked surface Ig attaches to actin. Nature. 1978 May 25;273(5660):278–281. doi: 10.1038/273278a0. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Hebdon M., Le Vine H., 3rd, Sahyoun N., Schmitges C. J., Cuatrecasas P. Properties of the interaction of fluoride- and guanylyl-5'-imidodiphosphate-regulatory proteins with adenylate cyclase. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3693–3697. doi: 10.1073/pnas.75.8.3693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ji T. H. The application of chemical crosslinking for studies on cell membranes and the identification of surface reporters. Biochim Biophys Acta. 1979 Apr 23;559(1):39–69. doi: 10.1016/0304-4157(79)90007-8. [DOI] [PubMed] [Google Scholar]
  16. Kennedy M. S., Insel P. A. Inhibitors of microtubule assembly enhance beta-adrenergic and prostaglandin E1-stimulated cyclic AMP accumulation in S49 lymphoma cells. Mol Pharmacol. 1979 Jul;16(1):215–223. [PubMed] [Google Scholar]
  17. Koch G. L., Smith M. J. An association between actin and the major histocompatibility antigen H-2. Nature. 1978 May 25;273(5660):274–278. doi: 10.1038/273274a0. [DOI] [PubMed] [Google Scholar]
  18. Lux S. E. Dissecting the red cell membrane skeleton. Nature. 1979 Oct 11;281(5731):426–429. doi: 10.1038/281426a0. [DOI] [PubMed] [Google Scholar]
  19. Madddy A. H., Kelly P. G. Acetic acid as a solvent for erythrocyte membrane proteins. Biochim Biophys Acta. 1971 Aug 13;241(2):290–301. doi: 10.1016/0005-2736(71)90029-0. [DOI] [PubMed] [Google Scholar]
  20. Marchesi V. T., Steers E., Jr Selective solubilization of a protein component of the red cell membrane. Science. 1968 Jan 12;159(3811):203–204. doi: 10.1126/science.159.3811.203. [DOI] [PubMed] [Google Scholar]
  21. Monneron A., d'Alayer J. Effects of crosslinking agents on adenylate cyclase regulation. FEBS Lett. 1980 Jan 1;109(1):75–80. doi: 10.1016/0014-5793(80)81315-9. [DOI] [PubMed] [Google Scholar]
  22. Orly J., Schramm M. Coupling of catecholamine receptor from one cell with adenylate cyclase from another cell by cell fusion. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4410–4414. doi: 10.1073/pnas.73.12.4410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pardoll D. M., Vogelstein B., Coffey D. S. A fixed site of DNA replication in eucaryotic cells. Cell. 1980 Feb;19(2):527–536. doi: 10.1016/0092-8674(80)90527-9. [DOI] [PubMed] [Google Scholar]
  24. Pearse B. M. On the structural and functional components of coated vesicles. J Mol Biol. 1978 Dec 25;126(4):803–812. doi: 10.1016/0022-2836(78)90021-9. [DOI] [PubMed] [Google Scholar]
  25. Rodbell M. The role of hormone receptors and GTP-regulatory proteins in membrane transduction. Nature. 1980 Mar 6;284(5751):17–22. doi: 10.1038/284017a0. [DOI] [PubMed] [Google Scholar]
  26. Sahyoun N., Cuatrecasas P. Mechanism of activation of adenylate cyclase by cholera toxin. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3438–3442. doi: 10.1073/pnas.72.9.3438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sahyoun N., Hollenberg M. D., Bennett V., Cuatrecasas P. Topographic separation of adenylate cyclase and hormone receptors in the plasma membrane of toad erythrocyte ghosts. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2860–2864. doi: 10.1073/pnas.74.7.2860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Singer S. J., Nicolson G. L. The fluid mosaic model of the structure of cell membranes. Science. 1972 Feb 18;175(4023):720–731. doi: 10.1126/science.175.4023.720. [DOI] [PubMed] [Google Scholar]
  29. Singer S. J. The molecular organization of membranes. Annu Rev Biochem. 1974;43(0):805–833. doi: 10.1146/annurev.bi.43.070174.004105. [DOI] [PubMed] [Google Scholar]
  30. Skerrow C. J. Selective extraction of desmosomal proteins by low ionic strength media. Biochim Biophys Acta. 1979 Jul 25;579(1):241–245. doi: 10.1016/0005-2795(79)90103-x. [DOI] [PubMed] [Google Scholar]
  31. Steck T. L., Weinstein R. S., Straus J. H., Wallach D. F. Inside-out red cell membrane vesicles: preparation and purification. Science. 1970 Apr 10;168(3928):255–257. doi: 10.1126/science.168.3928.255. [DOI] [PubMed] [Google Scholar]
  32. Steck T. L., Yu J. Selective solubilization of proteins from red blood cell membranes by protein perturbants. J Supramol Struct. 1973;1(3):220–232. doi: 10.1002/jss.400010307. [DOI] [PubMed] [Google Scholar]
  33. Sternweis P. C., Gilman A. G. Reconstitution of catecholamine-sensitive adenylate cyclase. Reconstitution of the uncoupled variant of the S40 lymphoma cell. J Biol Chem. 1979 May 10;254(9):3333–3340. [PubMed] [Google Scholar]
  34. Yu J., Fischman D. A., Steck T. L. Selective solubilization of proteins and phospholipids from red blood cell membranes by nonionic detergents. J Supramol Struct. 1973;1(3):233–248. doi: 10.1002/jss.400010308. [DOI] [PubMed] [Google Scholar]
  35. Zor U., Strulovici B., Lindner H. R. Implication of microtubules and microfilaments in the response of the ovarian adenylate cyclase-cyclic AMP system to gonadotropins and prostaglandin E2. Biochem Biophys Res Commun. 1978 Feb 28;80(4):983–992. doi: 10.1016/0006-291x(78)91342-6. [DOI] [PubMed] [Google Scholar]

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