Hostname: page-component-78c5997874-4rdpn Total loading time: 0 Render date: 2024-11-10T16:48:39.351Z Has data issue: false hasContentIssue false
  • English
  • Français

Photoreceptor rod outer segment 48-kDa protein has ATPase activity

Published online by Cambridge University Press:  02 June 2009

Ari Sitaramayya  and
Shereen Hakki
Ari Sitaramayya
Affiliation:
Department of Basic Sciences, Pennsylvania College of Optometry, Philadelphia
Shereen Hakki
Affiliation:
Department of Basic Sciences, Pennsylvania College of Optometry, Philadelphia
Get access Rights & Permissions [Opens in a new window]

Abstract

The role of 48-kDa protein in Visual transduction remains unresolved. Two hypotheses for its role in quenching the light activation of cyclic GMP cascade suggest that the protein binds to either phosphodiesterase or phosphorylated rhodopsin. Since the protein is also reported to bind ATP, we anticipated that the protein may have ATP hydrolyzing activity, and in analogy with the GTP-binding protein of the rod outer segments, such activity may be greatly enhanced by the elements of transduction cyclic GMP cascade, permitting the protein to function cyclically as GTP-binding protein does. We found that purified 48-kDa protein hydrolyzes ATP but at a slow rate of 0.04–0.05 per min. The Km for ATP is about 45–65 μM. The activity is inhibited noncompetitively by ADP with a Ki of about 50 μM. The ATPase activity of 48-kDa protein is not affected by rhodopsin, bleached rhodopsin, phosphorylated rhodopsin, unactivated cyclic GMP phosphodiesterase, or phosphodiesterase (PDE) activated by GMP PNP-bound G-protein. These data show that although 48-kDa protein has ATPase activity, lack of regulation of this activity by the elements of visual transduction makes it unlikely for this activity to have a role in quenching the light activation of cyclic GMP cascade.

Keywords

PhotoreceptorArrestinRhodopsinG-proteinPhosphodiesterase
Type
Research Article
Information
Visual Neuroscience , Volume 5 , Issue 6 , December 1990 , pp. 585 - 589
Copyright
Copyright © Cambridge University Press 1990

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Arshavsky, V. Yu., Dizhoor, A.M., Shestakova, I.K. & Philippov, P.P. (1985). The effect of rhodopsin phosphorylation on the light- dependent activation of phosphodiesterase from bovine rod outer segments. Federation of European Biochemical Societies Letters 181, 264–266.CrossRefGoogle ScholarPubMed
Baehr, W., Devlin, M.J. & Applebury, M.L. (1979). Isolation and characterization of cGMP phosphodiesterase from bovine rod outer segments. Journal of Biological Chemistry 254, 11669–11677.CrossRefGoogle ScholarPubMed
Baehr, W., Morita, E.A., Swanson, R.J. & Applebury, M.L. (1982). Characterization of bovine rod Outer segment G protein. Journal of Biological Chemistry 257, 6452–6460.CrossRefGoogle ScholarPubMed
Bennett, N. & Sitaramayya, A. (1988). Inactivation of photoexcited rhodopsin in retinal rods: the roles of rhodopsin kinase and 48-kDa protein (arrestin). Biochemistry 27, 1710–1715.CrossRefGoogle ScholarPubMed
Broekhuyse, R.M., Tolhuizen, E.F.J., Jaussen, A.P.M. & Winkens, H.J. (1985). The effect of light adaptation on the binding of 48-kDa protein (S antigen) to photoreceptor cell membranes. Current Eye Research 4, 613–618.CrossRefGoogle Scholar
Fung, B.K.-K. & Stryer, L. (1980). Photolyzed rhodopsin catalyzes the exchange of GTP for bound GDP in retinal rod outer segments. Proceedings of the National Academy of Sciences of the U.S.A. 77, 2500–2504.CrossRefGoogle Scholar
Glitscher, W. & Ruppel, H. (1989). Arrestin of bovine photoreceptors reveals strong ATP binding. Federation of European Biochemical Societies Letters 256, 101–105.CrossRefGoogle ScholarPubMed
Miller, J.L. & Dratz, E.A. (1984). Phosphorylation at sites near rhodopsin's carboxyl terminus regulates light-initiated cGMP hydrolysis. Vision Research 24, 1509–1521.CrossRefGoogle ScholarPubMed
Miller, J.L., Fox, D.A. & Litman, B.L. (1986). Amplification of phosphodiesterase activation is greatly reduced by rhodopsin phosphorylation. Biochemsitry 25, 4983–4988.CrossRefGoogle ScholarPubMed
Okada, D., Nakai, T. & Ikai, A. (1989). Transducin activation by molecular species of rhodopsin other than metarhodopsin, II: Photochemistry and Photobiology 49, 197–203.CrossRefGoogle ScholarPubMed
Pfister, C., Chabre, M., Plouet, J., Tuyen, V.V., Dekozak, Y., Faure, J.P. & Kuhn, H. (1985). Retinal S antigen identified as the 48k protein-regulating light-dependent phosphodiesterase in rods. Science 228, 891–893.CrossRefGoogle ScholarPubMed
Shichi, H., Somers, R.L. & Yamamoto, K. (1983). Rhodopsin kinase. Methods in Enzymology 99, 362–366.CrossRefGoogle ScholarPubMed
Shuster, T., Nagy, A.K. & Farber, D.B. (1988). Nucleotide binding to the rod outer segment rim protein. Experimental Eye Research 46, 647–655.CrossRefGoogle Scholar
Sitramayya, A. (1986). Rhodopsin kinase prepared from bovine rod disk membranes quenches light activation of cGMP phosphodiesterase in a reconstituted system. Biochemistry 25, 5460–5468.CrossRefGoogle Scholar
Sitaramayya, A., Casadevall, C., Bennett, N. & Hakki, S.I. (1988). Contribution of the guanosinetriphosphatase activity of G protein to termination of light-activated guanosine cyclic 3′,5′-phosphate hydrolysis in retinal rod outer segments. Biochemistry 27, 4880–4887.CrossRefGoogle Scholar
Thacher, S.M. (1982). Characterization of the light-activated Mg2+-ATPase in rod outer segments. Methods in Enzymology 81, 514–516.CrossRefGoogle ScholarPubMed
Uhl, R., Borys, T. & Abrahamson, E.W. (1982). Assays and characterization of Mg2+-ATPase in the rod outer segment of vertebrate photoreceptors. Methods in Enzymology 81, 509–513.CrossRefGoogle ScholarPubMed
Wheeler, G.L. & Bitensky, M.W. (1977). A light-activated GTPase in vertebrate photoreceptors: regulation of light-activated cyclic GMP phosphodiesterase. Proceedings of the National Academy of Sciences of the U.S.A. 74, 4238–4242.CrossRefGoogle ScholarPubMed
Wilden, U. & Kuhn, H. (1982). Light-dependent phosphorylation of rhodopsin: number of phosphorylation sites. Biochemsitry 21, 3014–3022.CrossRefGoogle ScholarPubMed
Wilden, U., Hall, S.W. & Kuhn, H. (1986 a). Phosphodiesterase activation by photoexcited rhodopsin is quenched when rhodopsin is phosphorylated and binds the intrinsic 48-kDa protein of rod outer segments. Proceedings of the National Academy of Sciences of the U.S.A. 83, 1174–1178.CrossRefGoogle ScholarPubMed
Wilden, U., Wust, E., Weyand, I. & Kuhn, H. (1986 b). Rapid affinity purification of retinal arrestin (48-kDa protein) via light-dependent binding to phosphorylated rhodopsin. Federation of European Biochemical Societies Letters 207, 292–295.CrossRefGoogle Scholar
Yamaki, K., Tsuda, M. & Shinohara, T. (1988). The sequence of human retinal S antigen reveals similarities with alpha-transducin. Federation of European Biochemical Societies Letters 234, 39–43.CrossRefGoogle ScholarPubMed
Zuckerman, R., Schmidt, G.J. & Dacko, S.M. (1982). Rhodopsin-tometarhodopsin II transition triggers amplified changes in cytosol ATP and ADP in intact retinal rod outer segments. Proceedings of the National Academy of Sciences of the U.S.A. 79, 6414–6418.CrossRefGoogle ScholarPubMed
Zuckerman, R., Buzdygon, B. & Liebman, P.A. (1984). Characterization of the 48-kDa protein of retinal rod outer segments as a light- dependent ATP-binding protein. Investigative Ophthalmology and Visual Science 25, 112.Google Scholar
Zuckerman, R., Buzdygon, B., Philp, N., Liebman, P. & Sitaramayya, A. (1985). Arrestin: an ATP/ADP exchange protein that regulates cGMP phosphodiesterase activity in retinal rod disk membrances (RDM). Biophysical Journal 47, 37a.Google Scholar
Zuckerman, R. & Cheasty, J.E. (1986). A 48-kDa protein arrests cGMP phosphodiesterase activation in retinal rod disk membranes. Federation of European Biochemical Societies Letters 207, 35–41.CrossRefGoogle Scholar