Skip to main content
PMC home page
The Journal of General Physiology logoLink to The Journal of General Physiology
. 1982 Apr 1;79(4):633–655. doi: 10.1085/jgp.79.4.633

Calcium and cyclic GMP regulation of light-sensitive protein phosphorylation in frog photoreceptor membranes

PMCID: PMC2215482  PMID: 6279759

Abstract

In frog photoreceptor membranes, light induces a dephosphorylation of two small proteins and a phosphorylation of rhodopsin. The level of phosphorylation of the two small proteins is influenced by cyclic GMP. Measurement of their phosphorylation as a function of cyclic GMP concentration shows fivefold stimulation as cyclic GMP is increased from 10(-5) to 10(-3) M. This includes the concentration range over which light activation of a cyclic GMP phosphodiesterase causes cyclic GMP levels to fall in vivo. Cyclic AMP does not affect the phosphorylations. Calcium ions inhibit the phosphorylation reactions. Calcium inhibits the cyclic GMP-stimulated phosphorylation of the small proteins as its concentration is increased from 10(-6) to 10(-3) M, with maximal inhibition of 70% being observed. Rhodopsin phosphorylation is not stimulated by cyclic nucleotides, but is inhibited by calcium, with 50% inhibition being observed as the Ca++ concentration is increased from 10(-9) to 10(-3) M. A nucleotide binding site appears to regulate rhodopsin phosphorylation. Several properties of the rhodopsin phosphorylation suggest that it does not play a role in a rapid ATP-dependent regulation of the cyclic GMP pathway. Calcium inhibition of protein phosphorylation is a distinctive feature of this system, and it is suggested that Ca++ regulation of protein phosphorylation plays a role in the visual adaptation process. Furthermore, the data provide support for the idea that calcium and cyclic GMP pathways interact in regulating the light-sensitive conductance.

Full Text

The Full Text of this article is available as a PDF (1.4 MB).

Selected References

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

  1. Baylor D. A., Hodgkin A. L. Changes in time scale and sensitivity in turtle photoreceptors. J Physiol. 1974 Nov;242(3):729–758. doi: 10.1113/jphysiol.1974.sp010732. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Biernbaum M. S., Bownds M. D. Influence of light and calcium on guanosine 5'-triphosphate in isolated frog rod outer segments. J Gen Physiol. 1979 Dec;74(6):649–669. doi: 10.1085/jgp.74.6.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bownds D., Dawes J., Miller J., Stahlman M. Phosphorylation of frog photoreceptor membranes induced by light. Nat New Biol. 1972 May 24;237(73):125–127. doi: 10.1038/newbio237125a0. [DOI] [PubMed] [Google Scholar]
  4. Bownds D., Gordon-Walker A., Gaide-Huguenin A. C., Robinson W. Characterization and analysis of frog photoreceptor membranes. J Gen Physiol. 1971 Sep;58(3):225–237. doi: 10.1085/jgp.58.3.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bownds M. D. Biochemical steps in visual transduction: roles for nucleotides and calcium ions. Photochem Photobiol. 1980 Oct;32(4):487–490. doi: 10.1111/j.1751-1097.1980.tb03792.x. [DOI] [PubMed] [Google Scholar]
  6. Brodie A. E., Bownds D. Biochemical correlates of adaptation processes in isolated frog photoreceptor membranes. J Gen Physiol. 1976 Jul;68(1):1–11. doi: 10.1085/jgp.68.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brown J. E., Coles J. A., Pinto L. H. Effects of injections of calcium and EGTA into the outer segments of retinal rods of Bufo marinus. J Physiol. 1977 Aug;269(3):707–722. doi: 10.1113/jphysiol.1977.sp011924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Castellucci V. F., Kandel E. R., Schwartz J. H., Wilson F. D., Nairn A. C., Greengard P. Intracellular injection of t he catalytic subunit of cyclic AMP-dependent protein kinase simulates facilitation of transmitter release underlying behavioral sensitization in Aplysia. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7492–7496. doi: 10.1073/pnas.77.12.7492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chader G. J., Fletcher R. T., O'Brien P. J., Krishna G. Differential phosphorylation by GTP and ATP in isolated rod outer segments of the retina. Biochemistry. 1976 Apr 20;15(8):1615–1620. doi: 10.1021/bi00653a004. [DOI] [PubMed] [Google Scholar]
  10. Chader G. J., Fletcher R. T., Russell P., Krishna G. Differential control of protein kinase activities of the retinal photoreceptor. Cation effects on phosphorylation by adenosine and guanosine 5'-triphosphates. Biochemistry. 1980 Jun 10;19(12):2634–2638. doi: 10.1021/bi00553a015. [DOI] [PubMed] [Google Scholar]
  11. Cohen A. I., Hall I. A., Ferrendelli J. A. Calcium and cyclic nucleotide regulation in incubated mouse retinas. J Gen Physiol. 1978 May;71(5):595–612. doi: 10.1085/jgp.71.5.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fain G. L. Sensitivity of toad rods: Dependence on wave-length and background illumination. J Physiol. 1976 Sep;261(1):71–101. doi: 10.1113/jphysiol.1976.sp011549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  14. Farber D. B., Brown B. M., Lllley R. N. Cyclic nucleotide dependent protein kinase and the phosphorylation of endogenous proteins of retinal rod outer segments. Biochemistry. 1979 Jan 23;18(2):370–378. doi: 10.1021/bi00569a022. [DOI] [PubMed] [Google Scholar]
  15. Fleischman D., Denisevich M. Guanylate cyclase of isolated bovine retinal rod axonemes. Biochemistry. 1979 Nov 13;18(23):5060–5066. doi: 10.1021/bi00590a006. [DOI] [PubMed] [Google Scholar]
  16. Frank R. N., Buzney S. M. Mechanism and specificity of rhodopsin phosphorylation. Biochemistry. 1975 Nov 18;14(23):5110–5117. doi: 10.1021/bi00694a014. [DOI] [PubMed] [Google Scholar]
  17. Frank R. N., Cavanagh H. D., Kenyon K. R. Light-stimulated phosphorylation of bovine visual pigments by adenosine triphosphate. J Biol Chem. 1973 Jan 25;248(2):596–609. [PubMed] [Google Scholar]
  18. Hagins W. A. The visual process: Excitatory mechanisms in the primary receptor cells. Annu Rev Biophys Bioeng. 1972;1:131–158. doi: 10.1146/annurev.bb.01.060172.001023. [DOI] [PubMed] [Google Scholar]
  19. Hagins W. A., Yoshikami S. Proceedings: A role for Ca2+ in excitation of retinal rods and cones. Exp Eye Res. 1974 Mar;18(3):299–305. doi: 10.1016/0014-4835(74)90157-2. [DOI] [PubMed] [Google Scholar]
  20. Hubbell W. L., Bownds M. D. Visual transduction in vertebrate photoreceptors. Annu Rev Neurosci. 1979;2:17–34. doi: 10.1146/annurev.ne.02.030179.000313. [DOI] [PubMed] [Google Scholar]
  21. Inouye M. Internal standards for molecular weight determinations of proteins by polyacrylamide gel electrophoresis. Applications to envelope proteins of Escherichia coli. J Biol Chem. 1971 Aug 10;246(15):4834–4838. [PubMed] [Google Scholar]
  22. Kaczmarek L. K., Jennings K. R., Strumwasser F., Nairn A. C., Walter U., Wilson F. D., Greengard P. Microinjection of catalytic subunit of cyclic AMP-dependent protein kinase enhances calcium action potentials of bag cell neurons in cell culture. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7487–7491. doi: 10.1073/pnas.77.12.7487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kawamura S., Bownds M. D. Light adaption of the cyclic GMP phosphodiesterase of frog photoreceptor membranes mediated by ATP and calcium ions. J Gen Physiol. 1981 May;77(5):571–591. doi: 10.1085/jgp.77.5.571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kleinschmidt J., Dowling J. E. Intracellular recordings from gecko photoreceptors during light and dark adaptation. J Gen Physiol. 1975 Nov;66(5):617–648. doi: 10.1085/jgp.66.5.617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Korenbrot J. I., Cone R. A. Dark ionic flux and the effects of light in isolated rod outer segments. J Gen Physiol. 1972 Jul;60(1):20–45. doi: 10.1085/jgp.60.1.20. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Krebs E. G., Beavo J. A. Phosphorylation-dephosphorylation of enzymes. Annu Rev Biochem. 1979;48:923–959. doi: 10.1146/annurev.bi.48.070179.004423. [DOI] [PubMed] [Google Scholar]
  27. Kuo J. F., Andersson R. G., Wise B. C., Mackerlova L., Salomonsson I., Brackett N. L., Katoh N., Shoji M., Wrenn R. W. Calcium-dependent protein kinase: widespread occurrence in various tissues and phyla of the animal kingdom and comparison of effects of phospholipid, calmodulin, and trifluoperazine. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7039–7043. doi: 10.1073/pnas.77.12.7039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kwok-Keung Fung B., Stryer L. Photolyzed rhodopsin catalyzes the exchange of GTP for bound GDP in retinal rod outer segments. Proc Natl Acad Sci U S A. 1980 May;77(5):2500–2504. doi: 10.1073/pnas.77.5.2500. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kühn H., Dreyer W. J. Light dependent phosphorylation of rhodopsin by ATP. FEBS Lett. 1972 Jan 15;20(1):1–6. doi: 10.1016/0014-5793(72)80002-4. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Levin R. M., Weiss B. Specificity of the binding of trifluoperazine to the calcium-dependent activator of phosphodiesterase and to a series of other calcium-binding proteins. Biochim Biophys Acta. 1978 May 3;540(2):197–204. doi: 10.1016/0304-4165(78)90132-0. [DOI] [PubMed] [Google Scholar]
  32. Liebman P. A., Pugh E. N., Jr ATP mediates rapid reversal of cyclic GMP phosphodiesterase activation in visual receptor membranes. Nature. 1980 Oct 23;287(5784):734–736. doi: 10.1038/287734a0. [DOI] [PubMed] [Google Scholar]
  33. Lipton S. A., Ostroy S. E., Dowling J. E. Electrical and adaptive properties of rod photoreceptors in Bufo marinus. I. Effects of altered extracellular Ca2+ levels. J Gen Physiol. 1977 Dec;70(6):747–770. doi: 10.1085/jgp.70.6.747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Lolley R. N., Brown B. M., Farber D. B. Protein phosphorylation in rod outer segments from bovine retina: cyclic nucleotide-activated protein kinase and its endogenous substrate. Biochem Biophys Res Commun. 1977 Sep 23;78(2):572–578. doi: 10.1016/0006-291x(77)90217-0. [DOI] [PubMed] [Google Scholar]
  35. McLaughlin S., Brown J. Diffusion of calcium ions in retinal rods. A theoretical calculation. J Gen Physiol. 1981 Apr;77(4):475–487. doi: 10.1085/jgp.77.4.475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Michaelson D. M., Avissar S. Ca2+-dependent protein phosphorylation of purely cholinergic Torpedo synaptosomes. J Biol Chem. 1979 Dec 25;254(24):12542–12546. [PubMed] [Google Scholar]
  37. Miller J. A., Paulsen R., Bownds M. D. Control of light-activated phosphorylation in frog photoreceptor membranes. Biochemistry. 1977 Jun 14;16(12):2633–2639. doi: 10.1021/bi00631a009. [DOI] [PubMed] [Google Scholar]
  38. O'Callaghan J. P., Dunn L. A., Lovenberg W. Calcium-regulated phosphorylation in synaptosomal cytosol: dependence on calmodulin. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5812–5816. doi: 10.1073/pnas.77.10.5812. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  40. Orr H. T., Lowry O. H., Cohen A. I., Ferrendelli J. A. Distribution of 3':5'-cyclic AMP and 3':5'-cyclic GMP in rabbit retina in vivo: selective effects of dark and light adaptation and ischemia. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4442–4445. doi: 10.1073/pnas.73.12.4442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Polans A. S., Hermolin J., Bownds M. D. Light-induced dephosphorylation of two proteins in frog rod outer segments: influence of cyclic nucleotides and calcium. J Gen Physiol. 1979 Nov;74(5):595–613. doi: 10.1085/jgp.74.5.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Polans A. S., Kawamura S., Bownds M. D. Influence of calcium on guanosine 3',5'-cyclic monophosphate levels in frog rod outer segments. J Gen Physiol. 1981 Jan;77(1):41–48. doi: 10.1085/jgp.77.1.41. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Robinson P. R., Kawamura S., Abramson B., Bownds M. D. Control of the cyclic GMP phosphodiesterase of frog photoreceptor membranes. J Gen Physiol. 1980 Nov;76(5):631–645. doi: 10.1085/jgp.76.5.631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Rudolph S. A., Krueger B. K. Endogenous protein phosphorylation and dephosphorylation. Adv Cyclic Nucleotide Res. 1979;10:107–133. [PubMed] [Google Scholar]
  45. Schoffeniels E., Dandrifosse G. Protein phosphorylation and sodium conductance in nerve membrane. Proc Natl Acad Sci U S A. 1980 Feb;77(2):812–816. doi: 10.1073/pnas.77.2.812. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Shichi H., Somers R. L. Light-dependent phosphorylation of rhodopsin. Purification and properties of rhodopsin kinase. J Biol Chem. 1978 Oct 10;253(19):7040–7046. [PubMed] [Google Scholar]
  47. Szuts E. Z. Calcium flux across disk membranes. Studies with intact rod photoreceptors and purified disks. J Gen Physiol. 1980 Sep;76(3):253–286. doi: 10.1085/jgp.76.3.253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Takai Y., Kishimoto A., Iwasa Y., Kawahara Y., Mori T., Nishizuka Y. Calcium-dependent activation of a multifunctional protein kinase by membrane phospholipids. J Biol Chem. 1979 May 25;254(10):3692–3695. [PubMed] [Google Scholar]
  49. Tillotson D., Gorman A. L. Non-uniform Ca2+ buffer distribution in a nerve cell body. Nature. 1980 Aug 21;286(5775):816–817. doi: 10.1038/286816a0. [DOI] [PubMed] [Google Scholar]
  50. Troyer E. W., Hall I. A., Ferrendelli J. A. Guanylate cyclases in CNS: enzymatic characteristics of soluble and particulate enzymes from mouse cerebellum and retina. J Neurochem. 1978 Oct;31(4):825–833. doi: 10.1111/j.1471-4159.1978.tb00117.x. [DOI] [PubMed] [Google Scholar]
  51. Wolff D. J., Brostrom C. O. Properties and functions of the calcium-dependent regulator protein. Adv Cyclic Nucleotide Res. 1979;11:27–88. [PubMed] [Google Scholar]
  52. Woodruff M. L., Bownds D., Green S. H., Morrisey J. L., Shedlovsky A. Guanosine 3',5'-cyclic monophosphate and the in vitro physiology of frog photoreceptor membranes. J Gen Physiol. 1977 May;69(5):667–679. doi: 10.1085/jgp.69.5.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Woodruff M. L., Bownds M. D. Amplitude, kinetics, and reversibility of a light-induced decrease in guanosine 3',5'-cyclic monophosphate in frog photoreceptor membranes. J Gen Physiol. 1979 May;73(5):629–653. doi: 10.1085/jgp.73.5.629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Wrenn R. W., Katoh N., Wise B. C., Kuo J. F. Stimulation by phosphatidylserine and calmodulin of calcium-dependent phosphorylation of endogenous proteins from cerebral cortex. J Biol Chem. 1980 Dec 25;255(24):12042–12046. [PubMed] [Google Scholar]
  55. Yau K. W., McNaughton P. A., Hodgkin A. L. Effect of ions on the light-sensitive current in retinal rods. Nature. 1981 Aug 6;292(5823):502–505. doi: 10.1038/292502a0. [DOI] [PubMed] [Google Scholar]
  56. Yee R., Liebman P. A. Light-activated phosphodiesterase of the rod outer segment. Kinetics and parameters of activation and deactivation. J Biol Chem. 1978 Dec 25;253(24):8902–8909. [PubMed] [Google Scholar]
  57. Yoshikami S., George J. S., Hagins W. A. Light-induced calcium fluxes from outer segment layer of vertebrate retinas. Nature. 1980 Jul 24;286(5771):395–398. doi: 10.1038/286395a0. [DOI] [PubMed] [Google Scholar]
  58. Yoshikami S., Robinson W. E., Hagins W. A. Topology of the outer segment membranes of retinal rods and cones revealed by a fluorescent probe. Science. 1974 Sep 27;185(4157):1176–1179. doi: 10.1126/science.185.4157.1176. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of General Physiology are provided here courtesy of The Rockefeller University Press

RESOURCES