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. 1978 Nov 1;175(2):421–430. doi: 10.1042/bj1750421

Topography of the rhodopsin molecule. Identification of the domain phosphorylated.

G J Sale, P Towner, M Akhtar
PMCID: PMC1186087  PMID: 743205

Abstract

Studies on the light-stimulated phosphorylation of rod outer segments by [gamma-32P]ATP showed that although nearly 1 mol of [32P]phosphate was incorporated/mol of total opsin, only a small fraction of the molecules were phosphorylated, and these contained at least 2-3 mol of phosphate/mol. Rod outer segments containing the phosphorylated opsin were incubated with 11-cis-retinal to generate phosphorylated rhodopsin and then digested with papain to produce a cleaved complex comprising three fragments, heavy (H), medium (M) and light (L). It was shown that L-fragment of apparent mol.wt. 6000 contained all the phosphorylation sites. This suggests that one specific domain of rhodopsin is susceptible to multiple phosphorylation.

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Selected References

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  1. Akhtar M., Blosse P. T., Dewhurst P. B. Studies on vision. The nature of the retinal-opsin linkage. Biochem J. 1968 Dec;110(4):693–702. doi: 10.1042/bj1100693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Akhtar M., Blosse P. T., Dewhurst P. B. The reduction of a rhodopsin derivative. Life Sci. 1965 Jun;4(12):1221–1226. doi: 10.1016/0024-3205(65)90336-x. [DOI] [PubMed] [Google Scholar]
  3. Applebury M. L., Zuckerman D. M., Lamola A. A., Jovin T. M. Rhodopsin. Purification and recombination with phospholipids assayed by the metarhodopsin I leads to metarhodopsin II transition. Biochemistry. 1974 Aug 13;13(17):3448–3458. doi: 10.1021/bi00714a005. [DOI] [PubMed] [Google Scholar]
  4. Bownds D., Brodie A., Robinson W. E., Palmer R. D., Miller J., Shedlovsky A. Proceedings: Physiology and enzymology of frog photoreceptor membranes. Exp Eye Res. 1974 Mar;18(3):253–269. doi: 10.1016/0014-4835(74)90153-5. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Bownds D. Site of attachment of retinal in rhodopsin. Nature. 1967 Dec 23;216(5121):1178–1181. doi: 10.1038/2161178a0. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Hargrave P. A. The amino-terminal tryptic peptide of bovine rhodopsin. A glycopeptide containing two sites of oligosaccharide attachment. Biochim Biophys Acta. 1977 May 27;492(1):83–94. doi: 10.1016/0005-2795(77)90216-1. [DOI] [PubMed] [Google Scholar]
  9. Hirtenstein M. D., Akhtar M. A convenient synthesis of labelled rhodopsin and studies on its active site. Biochem J. 1970 Sep;119(3):359–366. doi: 10.1042/bj1190359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kühn H., Cook J. H., Dreyer W. J. Phosphorylation of rhodopsin in bovine photoreceptor membranes. A dark reaction after illumination. Biochemistry. 1973 Jun 19;12(13):2495–2502. doi: 10.1021/bi00737a020. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Kühn H. Light-dependent phosphorylation of rhodopsin in living frogs. Nature. 1974 Aug 16;250(467):588–590. doi: 10.1038/250588a0. [DOI] [PubMed] [Google Scholar]
  13. MATHEWS C. K., BROWN F., COHEN S. S. VIRUS-INDUCED ACQUISITION OF METABOLIC FUNCTION. VII. BIOSYNTHESIS DE NOVO OF DEOXYCYTIDYLATE HYDROXYMETHYLASE. J Biol Chem. 1964 Sep;239:2957–2963. [PubMed] [Google Scholar]
  14. McDowell J. H., Kühn H. Light-induced phosphorylation of rhodopsin in cattle photoreceptor membranes: substrate activation and inactivation. Biochemistry. 1977 Sep 6;16(18):4054–4060. doi: 10.1021/bi00637a018. [DOI] [PubMed] [Google Scholar]
  15. Miller J. A., Paulsen R. Phosphorylation and dephosphorylation of frog rod outer segment membranes as part of the visual process. J Biol Chem. 1975 Jun 25;250(12):4427–4432. [PubMed] [Google Scholar]
  16. Röhlich P. Photoreceptor membrane carbohydrate on the intradiscal surface of retinal rod disks. Nature. 1976 Oct 28;263(5580):789–791. doi: 10.1038/263789a0. [DOI] [PubMed] [Google Scholar]
  17. Sale G. J., Towner P., Akhtar M. Functional rhodopsin complex consisting of three noncovalently linked fragments. Biochemistry. 1977 Dec 13;16(25):5641–5649. doi: 10.1021/bi00644a040. [DOI] [PubMed] [Google Scholar]
  18. Shichi H., Somers R. L., O'Brien P. J. Phosphorylation of rhodopsin: most rhodopsin molecules are not phosphorylated. Biochem Biophys Res Commun. 1974 Nov 6;61(1):217–221. doi: 10.1016/0006-291x(74)90555-5. [DOI] [PubMed] [Google Scholar]
  19. Swank R. T., Munkres K. D. Molecular weight analysis of oligopeptides by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate. Anal Biochem. 1971 Feb;39(2):462–477. doi: 10.1016/0003-2697(71)90436-2. [DOI] [PubMed] [Google Scholar]
  20. Towner P., Sale G. J., Akhtar M. Identification of the active site polypeptide in labeled photoreceptor membranes digested with papain. FEBS Lett. 1977 Apr 1;76(1):51–55. doi: 10.1016/0014-5793(77)80118-x. [DOI] [PubMed] [Google Scholar]
  21. Virmaux N., Weller M., Mandel P. Localisation of the major site. of light stimulated phosphorylation in a region of rhodopsin distinct from the chromophore binding site. FEBS Lett. 1975 May 15;53(3):320–323. doi: 10.1016/0014-5793(75)80046-9. [DOI] [PubMed] [Google Scholar]
  22. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  23. Weller M., Goridis C., Viramaux N., Mandel P. Letter: A hypothetical model for the possible involvement of rhodopsin phosphorylation in light and dark adaptation in the retina. Exp Eye Res. 1975 Oct;21(4):405–408. doi: 10.1016/0014-4835(75)90050-0. [DOI] [PubMed] [Google Scholar]
  24. Weller M., Virmaux N., Mandel P. Light-stimulated phosphorylation of rhodopsin in the retina: the presence of a protein kinase that is specific for photobleached rhodopsin. Proc Natl Acad Sci U S A. 1975 Jan;72(1):381–385. doi: 10.1073/pnas.72.1.381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Weller M., Virmaux N., Mandel P. Role of light and rhodopsin phosphorylation in control of permeability of retinal rod outer segment disks to Ca2plus. Nature. 1975 Jul 3;256(5512):68–70. doi: 10.1038/256068a0. [DOI] [PubMed] [Google Scholar]

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