Journal of Comparative Physiology A

, Volume 155, Issue 1, pp 39–45 | Cite as

Reversible phosphorylation of opsin induced by irradiation of blowfly retinae

  • R. Paulsen
  • J. Bentrop
Article

Summary

Light-induced phosphorylation and dephosphorylation of the visual pigment protein, opsin, was investigated in isolated retinae of the blowfly making use of the fact that photon capture by rhodopsin leads to the formation of a thermostable metarhodopsin. Retinae were exposed, in the presence of exogenous32P-orthophosphate, to an intense blue light which initiated the phosphorylation of opsin (half-time about 5 min at 25 °C). Subsequent exposure of the retina to red light converted all the metarhodopsin present into rhodopsin and triggered a relatively rapid dephosphorylation of rhodopsin (half-time less than 20 s). It is proposed that the phosphorylated forms of rhodopsin and metarhodopsin represent inactive states of the pigment, i.e. phosphorylated metarhodopsin does not initiate reactions leading to the excitation of the photoreceptor cell and phosphorylated rhodopsin cannot be converted into physiologically active metarhodopsin without first being dephosphorylated.

Keywords

Retina Blue Light Photoreceptor Cell Phosphorylated Form Inactive State 

Abbreviations

R1–6

peripheral retinula cells of the blowfly ommatidium

PDA

prolonged depolarizing afterpotential

R

rhodopsin

M

metarhodopsin

R-Pn

phosphorylated rhodopsin

M-Pn

phosphorylated metarhodopsin

SDS-PAGE

sodium dodecylsulphate polyacrylamide gel electrophoresis

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References

  1. Bownds D, Dawes J, Miller J, Stahlman M (1972) Phosphorylation of frog photoreceptor membranes induced by light. Nature 237:125–126Google Scholar
  2. Frank RN, Cavanagh HD, Kenyon KN (1973) Light-stimulated phosphorylation of bovine visual pigments by adenosine triphosphate. J Biol Chem 248:596–609Google Scholar
  3. Hamdorf K, Razmjoo S (1977) The prolonged depolarizing afterpotential and its contribution to the understanding of photoreceptor function. Biophys Struct Mech 3:163–170Google Scholar
  4. Hamdorf K, Razmjoo S (1979) Photoconvertible pigment states and excitation inCalliphora; the induction and properties of the prolonged depolarizing afterpotential. Biophys Struct Mech 5:137–161Google Scholar
  5. Hamdorf K, Paulsen R, Schwemer J (1973) Photoregeneration and sensitivity control of photoreceptors of invertebrates. In: Langer H (ed) Biochemistry and physiology of visual pigments. Springer, Berlin Heidelberg New York, pp 155–166Google Scholar
  6. Hillman P, Hochstein S, Minke B (1983) Transduction in invertebrate photoreceptors. Physiol Rev 63:668–772Google Scholar
  7. Kühn H (1974) Light-dependent phosphorylation of rhodopsin in living frogs. Nature 250:588–590Google Scholar
  8. Kühn H (1978) Light-regulated binding of rhodopsin kinase and other proteins to cattle photoreceptor membranes. Biochemistry 17:4389–4395Google Scholar
  9. Kühn H, Dreyer WJ (1972) Light dependent phosphorylation of rhodopsin by ATP. FEBS Lett 20:1–6Google Scholar
  10. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685Google Scholar
  11. Lee RH, Brown BM, Lolley RN (1981) Protein kinases of retinal rod outer segments: identification and partial characterization of cyclic nucleotide dependent protein kinase and rhodopsin kinase. Biochemistry 20:7532–7538Google Scholar
  12. Liebman PA, Pugh EN Jr (1980) ATP mediates rapid reversal of cyclic GMP phosphodiesterase activation in visual receptor membranes. Nature 287:734–736Google Scholar
  13. Matsumoto H, Pak WL (1984) Light-induced phosphorylation of retina-specific polypeptides ofDrosophila in vivo. Science 223:184–186Google Scholar
  14. Miller J, Paulsen R (1975) Phosphorylation and dephosphorylation of frog rod outer segment membranes as part of the visual process. J Biol Chem 250:4427–4432Google Scholar
  15. Miller J, Paulsen R, Bownds MD (1977) Control of light-activated phosphorylation in frog photoreceptor membranes. Biochemistry 16:2633–2639Google Scholar
  16. Minke B (1979) Transduction in photoreceptors with bistable pigments: intermediate processes. Biophys Struct Mech 5:163–174Google Scholar
  17. Oakley BR, Kirsch DR, Morris RM (1980) A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels. Anal Biochem 105:361–363Google Scholar
  18. Paulsen R (1984) Spectral characteristics of isolated blowfly rhabdoms. J Comp Physiol A 155:47–55Google Scholar
  19. Paulsen R, Bentrop J (1983) Activation of rhodopsin phosphorylation is triggered by the lumirhodopsin — metarhodopsin I transition. Nature 302:417–419Google Scholar
  20. Paulsen R, Hoppe I (1978) Light-activated phosphorylation of cephalopod rhodopsin. FEBS Lett 96:55–58Google Scholar
  21. Paulsen R, Schwemer J (1979) Vitamin A deficiency reduces the concentration of visual pigment protein within blowfly photoreceptor membranes. Biochim Biophys Acta 557:385–390Google Scholar
  22. Paulsen R, Schwemer J (1983) Biogenesis of blowfly photoreceptor membranes is regulated by 11-cis retinal. Europ J Biochem 137:609–614Google Scholar
  23. Paulsen R, Miller JA, Brodie AE, Bownds MD (1975) The decay of long-lived photoproducts in the isolated bull frog rod outer segment: relationship to other dark reactions. Vision Res 15:1325–1332Google Scholar
  24. Pfister C, Kühn H, Chabre M (1983) Interaction between photoexcited rhodopsin and peripheral enzymes in frog retinal rods. Eur J Biochem 136:489–199Google Scholar
  25. Razmjoo S, Hamdorf K (1980) In support of the ‘photopigment model’ of vision in invertebrates. J Comp Physiol 135:209–215Google Scholar
  26. Schwemer J (1983) Pathways of visual pigment regeneration in fly photoreceptor cells. Biophys Struct Mech 9:287–298Google Scholar
  27. Sitaramayya A, Liebman PA (1983) Phosphorylation of rhodopsin and quenching of cyclic GMP phosphodiesterase activation by ATP at weak bleaches. J Biol Chem 258:12106–12109Google Scholar
  28. Stavenga DG, Zantema A, Kuiper JW (1973) Rhodopsin processes and the function of the pupil mechanism in flies. In: Langer H (ed) Biochemistry and physiology of visual pigments. Springer, Berlin Heidelberg New York, pp 173–180Google Scholar

Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • R. Paulsen
    • 1
  • J. Bentrop
    • 1
  1. 1.Allgemeine Zoologie (Biologie I)Universität UlmUlmFederal Republic of Germany

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