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. 1990 Feb 1;265(3):655–658. doi: 10.1042/bj2650655

The effect of the gamma-subunit of the cyclic GMP phosphodiesterase of bovine and frog (Rana catesbiana) retinal rod outer segments on the kinetic parameters of the enzyme.

M M Whalen 1, M W Bitensky 1, D J Takemoto 1
PMCID: PMC1133684  PMID: 2154965

Abstract

Rod-outer-segment cyclic GMP phosphodiesterase (PDE) (subunit composition alpha beta gamma 2) contains catalytic activity in alpha beta. The gamma-subunits are inhibitors. Removal of the gamma-subunits increases Vmax. without affecting the Km. The inhibitory effect of a single gamma-subunit (alpha beta gamma) on the Vmax. of alpha beta is much greater in bovine than in frog (Rana catesbiana) PDE. Bovine PDE in the alpha beta gamma 2 state has a Vmax. that is 2.6 +/- 0.4% of the Vmax. of alpha beta. The removal of one gamma-subunit to give alpha beta gamma results in a Vmax. 5.2 +/- 1% of that for maximal activity. Frog alpha beta gamma 2 has a Vmax. 10.8 +/- 2%, and alpha beta gamma has a Vmax. 50 +/- 18%, of the Vmax. of alpha beta. These data suggest that a single gamma-subunit can inhibit the catalytic activity of active sites on both alpha- and beta-subunits in bovine, but not in frog, rod-outer-segment PDE.

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Selected References

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

  1. Baehr W., Devlin M. J., Applebury M. L. Isolation and characterization of cGMP phosphodiesterase from bovine rod outer segments. J Biol Chem. 1979 Nov 25;254(22):11669–11677. [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  3. Chabre M. Trigger and amplification mechanisms in visual phototransduction. Annu Rev Biophys Biophys Chem. 1985;14:331–360. doi: 10.1146/annurev.bb.14.060185.001555. [DOI] [PubMed] [Google Scholar]
  4. Cunnick J., Rider M., Takemoto L. J., Takemoto D. J. Rod/cone dysplasia in Irish setters. Presence of an altered rhodopsin. Biochem J. 1988 Mar 1;250(2):335–341. doi: 10.1042/bj2500335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Deterre P., Bigay J., Forquet F., Robert M., Chabre M. cGMP phosphodiesterase of retinal rods is regulated by two inhibitory subunits. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2424–2428. doi: 10.1073/pnas.85.8.2424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Deterre P., Bigay J., Robert M., Pfister C., Kühn H., Chabre M. Activation of retinal rod cyclic GMP-phosphodiesterase by transducin: characterization of the complex formed by phosphodiesterase inhibitor and transducin alpha-subunit. Proteins. 1986 Oct;1(2):188–193. doi: 10.1002/prot.340010210. [DOI] [PubMed] [Google Scholar]
  7. Deterre P., Bigay J., Robert M., Pfister C., Kühn H., Chabre M. Activation of retinal rod cyclic GMP-phosphodiesterase by transducin: characterization of the complex formed by phosphodiesterase inhibitor and transducin alpha-subunit. Proteins. 1986 Oct;1(2):188–193. doi: 10.1002/prot.340010210. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Godchaux W., 3rd, Zimmerman W. F. Membrane-dependent guanine nucleotide binding and GTPase activities of soluble protein from bovine rod cell outer segments. J Biol Chem. 1979 Aug 25;254(16):7874–7884. [PubMed] [Google Scholar]
  11. Halliday K. R., Stein P. J., Chernoff N., Wheeler G. L., Bitensky M. W. Limited trypsin proteolysis of photoreceptor GTP-binding protein. Light- and GTP-induced conformational changes. J Biol Chem. 1984 Jan 10;259(1):516–525. [PubMed] [Google Scholar]
  12. Hamm H. E., Bownds M. D. Protein complement of rod outer segments of frog retina. Biochemistry. 1986 Aug 12;25(16):4512–4523. doi: 10.1021/bi00364a010. [DOI] [PubMed] [Google Scholar]
  13. Hurley J. B., Stryer L. Purification and characterization of the gamma regulatory subunit of the cyclic GMP phosphodiesterase from retinal rod outer segments. J Biol Chem. 1982 Sep 25;257(18):11094–11099. [PubMed] [Google Scholar]
  14. Kohnken R. E., Eadie D. M., Revzin A., McConnell D. G. The light-activated GTP-dependent cyclic GMP phosphodiesterase complex of bovine retinal rod outer segments. Dark resolution of the catalytic and regulatory proteins. J Biol Chem. 1981 Dec 10;256(23):12502–12509. [PubMed] [Google Scholar]
  15. Kwok-Keung Fung B., Stryer L. Photolyzed rhodopsin catalyzes the exchange of GTP for bound GDP in retinal rod outer segments. Proc Natl Acad Sci U S A. 1980 May;77(5):2500–2504. doi: 10.1073/pnas.77.5.2500. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Miki N., Baraban J. M., Keirns J. J., Boyce J. J., Bitensky M. W. Purification and properties of the light-activated cyclic nucleotide phosphodiesterase of rod outer segments. J Biol Chem. 1975 Aug 25;250(16):6320–6327. [PubMed] [Google Scholar]
  18. Papermaster D. S. Preparation of retinal rod outer segments. Methods Enzymol. 1982;81:48–52. doi: 10.1016/s0076-6879(82)81010-0. [DOI] [PubMed] [Google Scholar]
  19. Sitaramayya A., Harkness J., Parkes J. H., Gonzalez-Oliva C., Liebman P. A. Kinetic studies suggest that light-activated cyclic GMP phosphodiesterase is a complex with G-protein subunits. Biochemistry. 1986 Feb 11;25(3):651–656. doi: 10.1021/bi00351a021. [DOI] [PubMed] [Google Scholar]
  20. Suter M. A modified ELISA technique for anti-hapten antibodies. J Immunol Methods. 1982 Aug 27;53(1):103–108. doi: 10.1016/0022-1759(82)90244-7. [DOI] [PubMed] [Google Scholar]
  21. Thompson W. J., Appleman M. M. Multiple cyclic nucleotide phosphodiesterase activities from rat brain. Biochemistry. 1971 Jan 19;10(2):311–316. [PubMed] [Google Scholar]
  22. Whalen M. M., Bitensky M. W. Comparison of the phosphodiesterase inhibitory subunit interactions of frog and bovine rod outer segments. Biochem J. 1989 Apr 1;259(1):13–19. doi: 10.1042/bj2590013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. 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]

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