Skip to main content
NCBI home page
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
. 1984 Aug;81(16):5111–5115. doi: 10.1073/pnas.81.16.5111

Squid rhodopsin and GTP-binding protein crossreact with vertebrate photoreceptor enzymes.

H R Saibil, M Michel-Villaz
PMCID: PMC391647  PMID: 6147847

Abstract

The activation of photoreceptor GTP-binding protein by rhodopsin was studied in squid photoreceptors and in crossreactions between the squid and bovine proteins. Turbidity changes were observed in the far-red after photoexcitation of rhodopsin with brief flashes and were used to probe interactions between photoreceptor membrane suspensions and soluble protein extracts. Our findings are squid photoreceptors contain a GTP-binding protein detectable by light- and GTP-sensitive turbidity changes and by limited sequence homology of a 46-kilodalton polypeptide to the alpha-subunit of bovine GTP-binding protein; the squid membranes activate bovine GTP-binding protein qualitatively in the same way as bovine rhodopsin; the 46-kilodalton component is present in a membrane-bound fraction but is more abundant in a crude, soluble fraction of squid rhabdomes, and this soluble fraction can interact with either squid or bovine rhodopsin-containing membranes; light-activated GTPase activities in all of these preparations are consistent with the light-induced turbidity changes. These results show that rhodopsin activation of GTP-binding protein is highly conserved in vertebrate and cephalopod photoreceptors. Since squid rhodopsin is immobilized in precisely ordered microvilli, this suggests that activation of GTP-binding protein in cephalopod photoreceptors occurs in the absence of rhodopsin diffusion. The rhodopsin immobility may be compensated by higher mobility of the soluble GTP-binding protein.

Full text

PDF
5111

Images in this article

5112Image
on p.5112
5114Image
on p.5114

Selected References

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

  1. Bennett N., Michel-Villaz M., Kühn H. Light-induced interaction between rhodopsin and the GTP-binding protein. Metarhodopsin II is the major photoproduct involved. Eur J Biochem. 1982 Sep;127(1):97–103. doi: 10.1111/j.1432-1033.1982.tb06842.x. [DOI] [PubMed] [Google Scholar]
  2. Bitensky M. W., Wheeler M. A., Rasenick M. M., Yamazaki A., Stein P. J., Halliday K. R., Wheeler G. L. Functional exchange of components between light-activated photoreceptor phosphodiesterase and hormone-activated adenylate cyclase systems. Proc Natl Acad Sci U S A. 1982 Jun;79(11):3408–3412. doi: 10.1073/pnas.79.11.3408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Calhoon R., Tsuda M., Ebrey T. G. A light-activated GTPase from octopus photoreceptors. Biochem Biophys Res Commun. 1980 Jun 30;94(4):1452–1457. doi: 10.1016/0006-291x(80)90582-3. [DOI] [PubMed] [Google Scholar]
  4. Calvert R., Gratzer W. B. Proteolytic digestion patterns of spectrin subunits. FEBS Lett. 1978 Feb 15;86(2):247–249. doi: 10.1016/0014-5793(78)80572-9. [DOI] [PubMed] [Google Scholar]
  5. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  6. Ebrey T., Tsuda M., Sassenrath G., West J. L., Waddell W. H. Light activation of bovine rod phosphodiesterase by non-physiological visual pigments. FEBS Lett. 1980 Jul 28;116(2):217–219. doi: 10.1016/0014-5793(80)80647-8. [DOI] [PubMed] [Google Scholar]
  7. Emeis D., Kühn H., Reichert J., Hofmann K. P. Complex formation between metarhodopsin II and GTP-binding protein in bovine photoreceptor membranes leads to a shift of the photoproduct equilibrium. FEBS Lett. 1982 Jun 21;143(1):29–34. doi: 10.1016/0014-5793(82)80266-4. [DOI] [PubMed] [Google Scholar]
  8. Fung B. K., Hurley J. B., Stryer L. Flow of information in the light-triggered cyclic nucleotide cascade of vision. Proc Natl Acad Sci U S A. 1981 Jan;78(1):152–156. doi: 10.1073/pnas.78.1.152. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hanski E., Rimon G., Levitzki A. Adenylate cyclase activation by the beta-adrenergic receptors as a diffusion-controlled process. Biochemistry. 1979 Mar 6;18(5):846–853. doi: 10.1021/bi00572a017. [DOI] [PubMed] [Google Scholar]
  10. Henis Y. I., Hekman M., Elson E. L., Helmreich E. J. Lateral motion of beta receptors in membranes of cultured liver cells. Proc Natl Acad Sci U S A. 1982 May;79(9):2907–2911. doi: 10.1073/pnas.79.9.2907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kühn H., Bennett N., Michel-Villaz M., Chabre M. Interactions between photoexcited rhodopsin and GTP-binding protein: kinetic and stoichiometric analyses from light-scattering changes. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6873–6877. doi: 10.1073/pnas.78.11.6873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kühn H. Light- and GTP-regulated interaction of GTPase and other proteins with bovine photoreceptor membranes. Nature. 1980 Feb 7;283(5747):587–589. doi: 10.1038/283587a0. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Liebman P. A., Mueller P., Pugh E. N., Jr Protons suppress the dark current of frog retinal rods. J Physiol. 1984 Feb;347:85–110. doi: 10.1113/jphysiol.1984.sp015055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Liebman P. A., Pugh E. N., Jr The control of phosphodiesterase in rod disk membranes: kinetics, possible mechanisms and significance for vision. Vision Res. 1979;19(4):375–380. doi: 10.1016/0042-6989(79)90097-x. [DOI] [PubMed] [Google Scholar]
  16. MacLeish P. R., Schwartz E. A., Tachibana M. Control of the generator current in solitary rods of the Ambystoma tigrinum retina. J Physiol. 1984 Mar;348:645–664. doi: 10.1113/jphysiol.1984.sp015131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Manning D. R., Gilman A. G. The regulatory components of adenylate cyclase and transducin. A family of structurally homologous guanine nucleotide-binding proteins. J Biol Chem. 1983 Jun 10;258(11):7059–7063. [PubMed] [Google Scholar]
  18. Miki N., Keirns J. J., Marcus F. R., Freeman J., Bitensky M. W. Regulation of cyclic nucleotide concentrations in photoreceptors: an ATP-dependent stimulation of cyclic nucleotide phosphodiesterase by light. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3820–3824. doi: 10.1073/pnas.70.12.3820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  20. Paulsen R., Zinkler D., Delmelle M. Architecture and dynamics of microvillar photoreceptor membranes of a cephalopod. Exp Eye Res. 1983 Jan;36(1):47–56. doi: 10.1016/0014-4835(83)90088-x. [DOI] [PubMed] [Google Scholar]
  21. Rodbell M., Lin M. C., Salomon Y., Londos C., Harwood J. P., Martin B. R., Rendell M., Berman M. Role of adenine and guanine nucleotides in the activity and response of adenylate cyclase systems to hormones: evidence for multisite transition states. Adv Cyclic Nucleotide Res. 1975;5:3–29. [PubMed] [Google Scholar]
  22. Romero-Saravia O., Hamdorf K. Alkaline phosphatase. A dominant enzyme of microvillus structure of cephalopod photoreceptors. Biochim Biophys Acta. 1983 Mar 23;729(1):90–96. doi: 10.1016/0005-2736(83)90459-5. [DOI] [PubMed] [Google Scholar]
  23. Saibil H. R. A light-stimulated increase of cyclic GMP in squid photoreceptors. FEBS Lett. 1984 Mar 26;168(2):213–216. doi: 10.1016/0014-5793(84)80248-3. [DOI] [PubMed] [Google Scholar]
  24. Saibil H. R. An ordered membrane-cytoskeleton network in squid photoreceptor microvilli. J Mol Biol. 1982 Jul 5;158(3):435–456. doi: 10.1016/0022-2836(82)90208-x. [DOI] [PubMed] [Google Scholar]
  25. Vandenberg C. A., Montal M. Light-regulated biochemical events in invertebrate photoreceptors. 1. Light-activated guanosinetriphosphatase, guanine nucleotide binding, and cholera toxin catalyzed labeling of squid photoreceptor membranes. Biochemistry. 1984 May 22;23(11):2339–2347. doi: 10.1021/bi00306a003. [DOI] [PubMed] [Google Scholar]
  26. Wheeler G. L., Bitensky M. W. A light-activated GTPase in vertebrate photoreceptors: regulation of light-activated cyclic GMP phosphodiesterase. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4238–4242. doi: 10.1073/pnas.74.10.4238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES