Advances in cyclic nucleotide and protein phosphorylation research

Many hormones and neurotransmitters exert their biological effects by increasing the levels of Ca2+ and 1,2-diacylglycerol in their target cells. Major agonists that act in this way are epinephrine and norepinephrine, acetylcholine, vasopressin, cholecystokinin, and angiotensin II. These and other Ca2+-mobilizing agonists may also produce effects that are not mediated by Ca2+ or diacylglycerol, but involve separate receptors and an increase or decrease in cyclic AMP. The general mechanisms by which Ca2+-mobilizing agonists induce their physiological responses are depicted in Fig. 12. These responses appear to involve an initial mobilization of Ca2+ from endoplasmic reticulum and perhaps other intracellular Ca2+ stores, followed by alterations in the flux of Ca2+ across the plasma membrane. The Ca2+ changes are consistently associated with increased turnover of cellular phosphoinositides. The most rapid response is breakdown of phosphatidylinositol 4,5-P2 in the plasma membrane, and there is much evidence that this involves a guanine-nucleotide-binding regulatory protein similar to those involved in the regulation of adenylate cyclase. Myo-inositol 1,4,5-P3 produced by phosphatidylinositol 4,5-P2 breakdown rapidly releases Ca2+ from endoplasmic reticulum, and it is likely that it is the long-sought second message for the Ca2+-dependent hormones. 1,2-Diacylglycerol, the other product of phosphatidylinositol 4,5-P2 breakdown, also acts as a second message in that it activates protein kinase C, a Ca2+-phospholipid-dependent protein kinase, by lowering its requirement for Ca2+. The cellular substrates for protein kinase C and its role in the different physiological responses to the Ca2+-mediated agonists are currently being defined. The major intracellular target for Ca2+ is the Ca2+-dependent regulatory protein calmodulin. This binds Ca2+ with high affinity, and the resulting complex interacts with a variety of enzymes and other cellular proteins, modifying their activities. A major target is the multifunctional calmodulin-dependent protein kinase that phosphorylates and alters the activities of many proteins, for example, glycogen synthase and tyrosine hydroxylase. Calcium ions may also stimulate calmodulin-dependent protein kinases that are more specific, such as phosphorylase kinase and myosin light-chain kinase. Other important Ca2+-calmodulin targets are the microtubule-associated proteins, but it is likely that many more will be found.(ABSTRACT TRUNCATED AT 400 WORDS)

FEBS LettersVolume 138, Issue 2 p. 157-163 Full-length articleFree Access Bimodal regulation of adenylate cyclase Dermot M.F. Cooper, Dermot M.F. Cooper Section on Membrane Regulation, Laboratory of Nutrition and Endocrinology, National Institute of Arthritis, Diabetes, Digestive and Kidney Diseases, National Institute of Health, Bethesda, MD 20205, USASearch for more papers by this author Dermot M.F. Cooper, Dermot M.F. Cooper Section on Membrane Regulation, Laboratory of Nutrition and Endocrinology, National Institute of Arthritis, Diabetes, Digestive and Kidney Diseases, National Institute of Health, Bethesda, MD 20205, USASearch for more papers by this author First published: February 22, 1982 https://doi.org/10.1016/0014-5793(82)80431-6Citations: 134AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat References 1 E.M. Ross, A.G. Gilman, Annu. Rev. Biochem., 49, (1980), 533– 564. 2 M. Rodbell, Nature, 284, (1980), 17– 22. 3 L.E. Limbird, Biochem. J., 195, (1981), 1– 13. 4 P.C. Sternweis, J.K. Northup, E. Hanski, L.S. Schleifer, M.D. Smigel, A.G. Gilman, Adv. Cyclic Nucl. Res., 14, (1981), 23– 36. 5 P.E. Cryer, L. Jarrett, D.M. Kipnis, Biochim. Biophys. Acta, 177, (1969), 586– 590. 6 J.P. Harwood, H. Low, M. Rodbell, J. Biol. Chem., 248, (1973), 6239– 6245. 7 R. Ebert, U. Schwabe, Naunyn-Schmiedebergs Arch. Pharmacol., 286, (1974), 297– 313. 8 J.N. Fain, R.H. Pointer, W.F. Ward, J. Biol. Chem., 247, (1972), 6866– 6872. 9 J. Moskowitz, J.P. Harwood, W.D. Reid, G. Krishna, Biochim. Biophys. Acta, 230, (1971), 279– 285. 10 F. Murad, Y.-M. Chi, T.W. Rall, E.W. Sutherland, J. Biol. Chem., 237, (1962), 1233– 1238. 11 H.O.J. Collier, A.C. Roy, Nature, 248, (1974), 24– 27. 12 K.H. Jakobs, W. Saur, G. Schultz, FEBS Lett., 85, (1978), 167– 170. 13 S.L. Sabol, M. Nirenberg, J. Biol. Chem., 254, (1979), 1913– 1920. 14 C. Londos, D.M.F. Cooper, W. Schlegel, M. Rodbell, Proc. Natl. Acad. Sci. USA, 75, (1978), 5362– 5366. 15 H. Yamamura, P.M. Lad, M. Rodbell, J. Biol. Chem., 252, (1977), 7964– 7966. 16 D.M.F. Cooper, W. Schlegel, M.C. Lin, M. Rodbell, J. Biol. Chem., 254, (1979), 8927– 8930. 17 A.J. Blume, D. Lichtshtein, G. Boone, Proc. Natl. Acad. Sci. USA, 76, (1979), 5626– 5630. 18 D. Lichtshtein, G. Boone, A.J. Blume, J. Cyclic Nucl. Res., 5, (1979), 367– 375. 19 K.H. Jakobs, K. Aktories, G. Schultz, Naunyn-Schmiedebergs Arch. Pharmacol., 310, (1979), 113– 119. 20 A.M. Watanabe, M.M. McConnaughey, R.A. Strawbridge, J.W. Fleming, L.R. Jones, H.R. Besch jr, J. Biol. Chem., 253, (1978), 4833– 4836. 21 T.E. Cote, C.W. Grewe, J.W. Kebabian, Endocrinology, 108, (1981), 420– 426. 22 C. Londos, D.M.F. Cooper, M. Rodbell, Adv. Cyclic Nucl. Res., 14, (1981), 163– 171. 23 R.J. Schimmel, K.K. McMahon, R. Serio, Mol. Pharmacol., 19, (1981), 248– 255. 24 K. Aktories, K.H. Jakobs, G. Schultz, FEBS Lett., 115, (1980), 11– 14. 25 D.M.F. Cooper, M. Rodbell, Nature, 282, (1979), 517– 518. 26 J.-M. Girardot, D.M.F. Cooper, J. Kempf, Adv. Cyclic Nucl. Res., 14, (1981), 657– 27 P.Y. Law, J. Wu, J.E. Koehler, H.H. Loh, J. Neurochem., 36, (1981), 1834– 1846. 28 D.M.F. Cooper, C. Londos, D.L. Gill, M. Rodbell, J. Neurochem., (1982), in press 29 S. Jard, B. Cantau, K.H. Jakobs, J. Biol. Chem., 256, (1981), 2603– 2606. 30 D.M.F. Cooper, C. Londos, M. Rodbell, Mol. Pharmacol., 18, (1980), 598– 601. 31 D.M.F. Cooper, C. Londos, L.D. Kohn Horizons in Biochemistry and Biophysics 6, (1982), Wiley London 309– 333. 32 K.H. Jakobs, Mol. Cell Endocrinol., 16, (1979), 147– 156. 33 C.B. Pert, S.H. Snyder, Mol. Pharmacol., 10, (1974), 868– 879. 34 B.S. Tsai, R.J. Lefkowitz, Mol. Pharmacol., 16, (1979), 61– 68. 35 T. Michel, B.B. Hoffman, R.J. Lefkowitz, Nature, 288, (1980), 709– 711. 36 A.J. Blume, Proc. Natl. Acad. Sci. USA, 75, (1978), 1713– 1717. 37 D.A. Greenberg, D.C. U'Prichard, P. Sheehan, S.H. Snyder, Brain Res., 140, (1978), 378– 384. 38 D.C. U'Prichard, S.H. Snyder, J. Neurochem., 34, (1980), 385– 394. 39 P.V. Sulakhe, N.L.-K Leung, A.T. Arbus, S.J. Sulakhe, S.H. Jan, N. Narayanan, Biochem. J., 164, (1977), 67– 74. 40 D. Evain, W.B. Anderson, J. Biol. Chem., 254, (1979), 8726– 8729. 41 F. Propst, B. Hamprecht, J. Neurochem., 36, (1981), 580– 588. 42 K.H. Jakobs, G. Schultz, Naunyn-Schmiedebergs Arch. Pharmacol., 310, (1979), 121– 127. 43 G. Koski, W.A. Klee, Proc. Natl. Acad. Sci. USA, 78, (1981), 4185– 4189. 44 K. Aktories, K.H. Jakobs, FEBS Lett., 130, (1981), 235– 238. 45 M.D. Resh, R.A. Nemenoff, G. Guidotti, J. Biol. Chem., 255, (1980), 10938– 10945. 46 M.A. Titheradge, J.L. Stringer, R.C. Haynes jr, Eur. J. Biochem., 102, (1979), 117– 124. 47 W.F. Simonds, G. Koski, R.A. Streaty, L.M. Hjelmeland, W.A. Klee, Proc. Natl. Acad. Sci. USA, 77, (1980), 4623– 4627. 48 U.T. Ruegg, S. Cuenod, J.M. Hiller, T. Gioannini, R.D. Howells, E.J. Simon, Proc. Natl. Acad. Sci. USA, 78, (1981), 4635– 4638. 49 G. Guellaen, M. Aggerbeck, J. Hanoune, J. Biol. Chem., 254, (1979), 10761– 10768. 50 S.K. Smith, L.E. Limbird, Proc. Natl. Acad. Sci. USA, 78, (1981), 4026– 4030. 51 D.M.F. Cooper, R. Jagus, R.L. Somers, M. Rodbell, Biochem. Biophys. Res. Commun., 101, (1981), 1179– 1185. 52 E. Hazum, K.J. Chang, P. Cuatrecasas, Science, 206, (1979), 1077– 1079. 53 H.B. Peng, P.C. Cheng, P.W. Luther, Nature, 292, (1981), 831– 834. 54 W. Schlegel, D.M.F. Cooper, M. Rodbell, Arch. Biochem. Biophys., 201, (1980), 678– 682. 55 J. Orly, M. Schramm, Proc. Natl. Acad. Sci. USA, 73, (1976), 4410– 4414. 56 H. Glossmann, A. Baukal, K.J. Catt, J. Biol. Chem., 249, (1974), 664– 666. 57 L.M. Jones, R.H. Michell, Biochem. Soc. Trans., 6, (1978), 673– 693. 58 S.K. Sharma, W.A. Klee, M. Nirenberg, Proc. Natl. Acad. Sci. USA, 72, (1975), 3092– 3096. 59 J. Finidori-Lepicard, S. Schorderet-Slatkine, J. Hanoune, E.E. Baulieu, Nature, 292, (1981), 255– 257. 60 S.E. Sadler, J.L. Maller, J. Biol. Chem., 256, (1981), 6368– 6373. 61 H. Liao, J. Thorner, Proc. Natl. Acad. Sci. USA, 77, (1980), 1898– 1902. Citing Literature Volume138, Issue2February 22, 1982Pages 157-163 ReferencesRelatedInformation

The attenuation of cyclic AMP accumulation occurs by different mechanisms in 1321N1 astrocytoma cells and NG108-15 neuroblastoma X glioma cells. In 1321N1 cells, cholinergic agonists reduce cyclic AMP accumulation through a Ca2+-dependent activation of phosphodiesterase; in NG108-15 cells, muscarinic receptor-mediated effects on cyclic AMP metabolism occur through inhibition of adenylate cyclase. The goal of the current study was to determine whether different pharmacological specificities were expressed by the muscarinic receptor populations of these two cell lines. The affinity of muscarinic receptors for [3H]quinuclidinyl benzilate (6 pM), [3H]N-methylscopolamine (50 pM), and atropine (80 pM) was similar in membrane preparations from each cell line. The affinity of the antagonist, pirenzepine, which has been proposed to be a selective ligand for a muscarinic receptor subtype, was 3-fold higher in competition binding assays carried out with membranes of 1321N1 cells, than with NG108-15 cells. The Hill coefficients of pirenzepine competition curves were not significantly different from unity in both cell lines. This selectivity of pirenzepine was also apparent in studies of the competitive inhibition of carbachol-induced attenuation of cyclic AMP accumulation in intact cells. Differences in the relative affinities of agonists were observed in competition binding analyses carried out with membranes in the presence of GTP and absence of Mg2+. The Ki values of bethanechol and carbachol were 5- and 12-fold lower for receptors of NG108-15 cells than those of 1321N1 cells and the Ki of methacholine was 3.5-fold lower for 1321N1 cells than for NG108-15 cells. The affinities of oxotremorine and arecoline were similar between the two cell lines. These differences in agonist affinities between the two cell lines were much smaller in analyses of muscarinic receptor-mediated effects on cyclic AMP metabolism in intact cells. Taken together, these data suggest that muscarinic receptors of differing pharmacological specificities regulate cyclic AMP metabolism by different mechanisms in 1321N1 and NG108-15 cells.

Progesterone inhibits oocyte plasma membrane adenylate cyclase measured in the presence of GTP or Gpp(NH)p by a novel mechanism that involves a guanine nucleotide regulatory protein. A hormone receptor has been identified in the oocyte plasma membranes using the technique of photoaffinity labeling, and the amount of steroid covalently bound to the steroid receptor after photolysis correlates with the level of inhibition of adenylate cyclase activity and the EC50 for germinal vesicle breakdown. Inhibition of oocyte adenylate cyclase by both progesterone and 2', 5'-dideoxyadenosine, a potent P-site agonist, correlates with slowing of guanine nucleotide exchange. The steroid inhibition shares certain other common characteristics with P-site action, including inhibition of Gpp(NH)p-stimulated enzyme activity and a slowing of the rate of Gpp(NH)p activation of the enzyme that is inversely proportional to the concentration of guanine nucleotide. The steady-state velocity of the activated enzyme is also reduced by both hormones. However, a major difference between the actions of progesterone and the P-site agonist is in the effects of the divalent cation Mn2+. Whereas Mn2+ potentiates the inhibitory action of 2', 5'-DDA, the divalent cation abolishes the inhibitory action of progesterone, as would be predicted for receptor-mediated action. The lack of effect of IAP on progesterone inhibition of oocyte adenylate cyclase suggests that progesterone inhibition of oocyte adenylate cyclase is not mediated by the IAP substrate. Possible alternative models for the IAP-insensitive steroid inhibition of oocyte adenylate cyclase include a unique interaction with Ni that is not abolished by IAP or an action that involves Ns.

High-affinity forskolin binding sites in brain membranes have been identified that have structure-activity characteristics compatible with forskolin's site of action at the adenylate cyclase enzyme. It is proposed that these high-affinity binding sites are associated with an activated complex of the catalytic protein and the alpha s subunit. Quantitation of high-affinity forskolin binding sites may provide a direct measure of the amount of adenylate cyclase that has the potential to be regulated by stimulatory hormones and the Ns subunit.