Abstract
An inhibitory subunit (P gamma) of cGMP phosphodiesterase from vertebrate rod photoreceptors (frog, toad, and bovine) was phosphorylated by cytosolic protein kinase(s) derived from intact frog rod outer segments. The phosphorylation of frog P gamma was stimulated by phosphatidylinositol but not by cAMP or cGMP. One- and two-dimensional gel electrophoresis revealed that 70-80% of P gamma was phosphorylated with 1 mol of phosphate per frog P gamma under optimal conditions. A peptide that derived from an active domain of bovine P gamma was also phosphorylated. Phosphorylation of frog P gamma was inhibited by addition of the peptide to the reaction mixture. Phosphorylation of frog P gamma was also inhibited by addition of transducin subunits or active (P gamma-less) cGMP phosphodiesterase. Okadaic acid, on the other hand, enhanced P gamma phosphorylation, suggesting the presence of protein phosphatase(s) in the cytosolic fraction. These data suggest another mechanism for the regulation of cGMP phosphodiesterase in vertebrate rod photoreceptors.
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Applebury M. L., Hargrave P. A. Molecular biology of the visual pigments. Vision Res. 1986;26(12):1881–1895. doi: 10.1016/0042-6989(86)90115-x. [DOI] [PubMed] [Google Scholar]
- Binder B. M., Brewer E., Bownds M. D. Stimulation of protein phosphorylations in frog rod outer segments by protein kinase activators. Suppression of light-induced changes in membrane current and cGMP by protein kinase C activators. J Biol Chem. 1989 May 25;264(15):8857–8864. [PubMed] [Google Scholar]
- Brown J. E., Blazynski C., Cohen A. I. Light induces a rapid and transient increase in inositol-trisphosphate in toad rod outer segments. Biochem Biophys Res Commun. 1987 Aug 14;146(3):1392–1396. doi: 10.1016/0006-291x(87)90804-7. [DOI] [PubMed] [Google Scholar]
- Cohen P., Klumpp S., Schelling D. L. An improved procedure for identifying and quantitating protein phosphatases in mammalian tissues. FEBS Lett. 1989 Jul 3;250(2):596–600. doi: 10.1016/0014-5793(89)80803-8. [DOI] [PubMed] [Google Scholar]
- Fesenko E. E., Kolesnikov S. S., Lyubarsky A. L. Induction by cyclic GMP of cationic conductance in plasma membrane of retinal rod outer segment. Nature. 1985 Jan 24;313(6000):310–313. doi: 10.1038/313310a0. [DOI] [PubMed] [Google Scholar]
- Fowles C., Akhtar M., Cohen P. Interplay of phosphorylation and dephosphorylation in vision: protein phosphatases of bovine rod outer segments. Biochemistry. 1989 Nov 28;28(24):9385–9391. doi: 10.1021/bi00450a020. [DOI] [PubMed] [Google Scholar]
- Gehm B. D., Mc Connell D. G. Phosphoinositide synthesis in bovine rod outer segments. Biochemistry. 1990 Jun 12;29(23):5442–5446. doi: 10.1021/bi00475a005. [DOI] [PubMed] [Google Scholar]
- Ghalayini A., Anderson R. E. Phosphatidylinositol 4,5-bisphosphate: light-mediated breakdown in the vertebrate retina. Biochem Biophys Res Commun. 1984 Oct 30;124(2):503–506. doi: 10.1016/0006-291x(84)91582-1. [DOI] [PubMed] [Google Scholar]
- Hamm H. Regulation by light of cyclic nucleotide-dependent protein kinases and their substrates in frog rod outer segments. J Gen Physiol. 1990 Mar;95(3):545–567. doi: 10.1085/jgp.95.3.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hayashi F., Amakawa T. Light-mediated breakdown of phosphatidylinositol-4,5-bisphosphate in isolated rod outer segments of frog photoreceptor. Biochem Biophys Res Commun. 1985 Apr 30;128(2):954–959. doi: 10.1016/0006-291x(85)90139-1. [DOI] [PubMed] [Google Scholar]
- Hayashi F., Sumi M., Amakawa T. Phosphatidylinositol stimulates phosphorylation of protein components I and II in rod outer segments of frog photoreceptors. Biochem Biophys Res Commun. 1987 Oct 14;148(1):54–60. doi: 10.1016/0006-291x(87)91075-8. [DOI] [PubMed] [Google Scholar]
- Ho Y. K., Hingorani V. N., Navon S. E., Fung B. K. Transducin: a signaling switch regulated by guanine nucleotides. Curr Top Cell Regul. 1989;30:171–202. doi: 10.1016/b978-0-12-152830-0.50008-6. [DOI] [PubMed] [Google Scholar]
- Lee R. H., Brown B. M., Lolley R. N. Autophosphorylation of rhodopsin kinase from retinal rod outer segments. Biochemistry. 1982 Jul 6;21(14):3303–3307. doi: 10.1021/bi00257a009. [DOI] [PubMed] [Google Scholar]
- Lee R. H., Brown B. M., Lolley R. N. Light-induced dephosphorylation of a 33K protein in rod outer segments of rat retina. Biochemistry. 1984 Apr 24;23(9):1972–1977. doi: 10.1021/bi00304a014. [DOI] [PubMed] [Google Scholar]
- Liebman P. A., Parker K. R., Dratz E. A. The molecular mechanism of visual excitation and its relation to the structure and composition of the rod outer segment. Annu Rev Physiol. 1987;49:765–791. doi: 10.1146/annurev.ph.49.030187.004001. [DOI] [PubMed] [Google Scholar]
- Morrison D. F., Rider M. A., Takemoto D. J. Modulation of retinal transducin and phosphodiesterase activities by synthetic peptides of the phosphodiesterase gamma-subunit. FEBS Lett. 1987 Oct 5;222(2):266–270. doi: 10.1016/0014-5793(87)80383-6. [DOI] [PubMed] [Google Scholar]
- Nagao S., Yamazaki A., Bitensky M. W. Calmodulin and calmodulin binding proteins in amphibian rod outer segments. Biochemistry. 1987 Mar 24;26(6):1659–1665. doi: 10.1021/bi00380a026. [DOI] [PubMed] [Google Scholar]
- Ovchinnikov YuA, Lipkin V. M., Kumarev V. P., Gubanov V. V., Khramtsov N. V., Akhmedov N. B., Zagranichny V. E., Muradov K. G. Cyclic GMP phosphodiesterase from cattle retina. Amino acid sequence of the gamma-subunit and nucleotide sequence of the corresponding cDNA. FEBS Lett. 1986 Aug 18;204(2):288–292. doi: 10.1016/0014-5793(86)80830-4. [DOI] [PubMed] [Google Scholar]
- Palczewski K., Hargrave P. A., McDowell J. H., Ingebritsen T. S. The catalytic subunit of phosphatase 2A dephosphorylates phosphoopsin. Biochemistry. 1989 Jan 24;28(2):415–419. doi: 10.1021/bi00428a001. [DOI] [PubMed] [Google Scholar]
- 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]
- Shinozawa T., Yoshizawa T. Cyclic nucleotide-dependent phosphorylation of proteins in rod outer segments in frog retina. Characteristics of the phosphorylated proteins and their dephosphorylation. J Biol Chem. 1986 Jan 5;261(1):216–223. [PubMed] [Google Scholar]
- Stryer L., Bourne H. R. G proteins: a family of signal transducers. Annu Rev Cell Biol. 1986;2:391–419. doi: 10.1146/annurev.cb.02.110186.002135. [DOI] [PubMed] [Google Scholar]
- Tuteja N., Farber D. B. Gamma-subunit of mouse retinal cyclic-GMP phosphodiesterase: cDNA and corresponding amino acid sequence. FEBS Lett. 1988 May 9;232(1):182–186. doi: 10.1016/0014-5793(88)80413-7. [DOI] [PubMed] [Google Scholar]
- Yamada T., Takeuchi Y., Komori N., Kobayashi H., Sakai Y., Hotta Y., Matsumoto H. A 49-kilodalton phosphoprotein in the Drosophila photoreceptor is an arrestin homolog. Science. 1990 Apr 27;248(4954):483–486. doi: 10.1126/science.2158671. [DOI] [PubMed] [Google Scholar]
- Yamazaki A., Bartucca F., Ting A., Bitensky M. W. Reciprocal effects of an inhibitory factor on catalytic activity and noncatalytic cGMP binding sites of rod phosphodiesterase. Proc Natl Acad Sci U S A. 1982 Jun;79(12):3702–3706. doi: 10.1073/pnas.79.12.3702. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yamazaki A., Hayashi F., Tatsumi M., Bitensky M. W., George J. S. Interactions between the subunits of transducin and cyclic GMP phosphodiesterase in Rana catesbiana rod photoreceptors. J Biol Chem. 1990 Jul 15;265(20):11539–11548. [PubMed] [Google Scholar]
- Yamazaki A., Stein P. J., Chernoff N., Bitensky M. W. Activation mechanism of rod outer segment cyclic GMP phosphodiesterase. Release of inhibitor by the GTP/GTP-binding protein. J Biol Chem. 1983 Jul 10;258(13):8188–8194. [PubMed] [Google Scholar]
- Yamazaki A., Tatsumi M., Bitensky M. W. Purification of rod outer segment GTP-binding protein subunits and cGMP phosphodiesterase by single-step column chromatography. Methods Enzymol. 1988;159:702–710. doi: 10.1016/0076-6879(88)59065-1. [DOI] [PubMed] [Google Scholar]
- Yamazaki A., Tatsumi M., Torney D. C., Bitensky M. W. The GTP-binding protein of rod outer segments. I. Role of each subunit in the GTP hydrolytic cycle. J Biol Chem. 1987 Jul 5;262(19):9316–9323. [PubMed] [Google Scholar]