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Site-directed mutagenesis of the cGMP phosphodiesterase inhibitory γ subunit from bovine rods

New mechanisms of catalytic subunits inhibition and activation

  • Chapter
Plant Hormone Signal Perception and Transduction

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Abstract

The recombinant and 30 mutant cGMP phosphodiesterase (PDE) γ subunit (PDEγ) genes were expressed by sequential transcription and translation in vitro. Inhibitory properties of these mutants and their interactions with PDE catalytic and transducin a subunits were studied. To explain the properties of PDEγ mutants we proposed a new mechanism of PDE functioning: (i) There are two sites on the PDE catalytic complex (PDEαβ) for PDEγ binding (A and B). PDEγ sites interacting with sites A and B are structurally different. The site on PDEγ that interacts with the B-site partly overlaps with the binding site for transducin α subunit complexed with GTP (Tα* GTP). (ii) The PDEγ bound to the B-site provides the main contribution to inhibition of the PDE catalytic activity, (iii) Tα* GTP first interacts with the PDEγ bound to the A-site and removes it in a PDEγ*(Tα*GTP) complex, (iv) After removal of PDEγ from the A-site, another Tα* GTP molecule is enabled to interact both with PDEαβ and with PDEγ bound to the B-site. This interaction results in the formation of a membrane-bound fully catalytically active triple complex, PDEαβ*PDEγ*(Tα*GTP).

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References

  1. Artemyev NO, Rarick HM, Mills JS, Skiba NP and Hamm HE (1992) Sites of interaction between rod G-protein α-subunit and cGMP-phosphodiesterase γ-subunit. J Biol Chem 267: 25067–25072

    PubMed  CAS  Google Scholar 

  2. Brown RL (1992) Functional regions of the inhibitory subunit of retinal rod cGMP phosphodiesterase identified by site-specific mutagenesis and fluorescent spectroscopy. Biochemistry 31: 5918–5925

    Article  PubMed  CAS  Google Scholar 

  3. Clerc A and Bennett N (1992) Activated cGMP phosphodiesterase of retinal rods. J Biol Chem 267: 6620–6627

    PubMed  CAS  Google Scholar 

  4. Cunnick JM, Hurt D, Oppert B, Sakamoto K and Takemoto DJ (1990) Binding of the γ-subunit of retinal rod-outer-segment phosphodiesterase with both transducin and the catalytic subunits of phosphodiesterase. Biochem J 271: 721–727

    PubMed  CAS  Google Scholar 

  5. Fung BK-K, Hurley JB and Stryer L (1981) Flow of information in the light-triggered cyclic nucleotide cascade of vision. Proc Natl Acad Sci USA 78: 152–156

    Article  PubMed  CAS  Google Scholar 

  6. Eibl H and Lands WEM (1969) Sensitive determination of phosphate. Anal Biochem 30: 51–57

    Article  PubMed  CAS  Google Scholar 

  7. Hurley JB and Stryer L (1982) Purification and characterization of the γ regulatory subunit of the cyclic GMP phospho-diesterase from retinal rod outer segments. J Biol Chem 257: 11094–11099

    PubMed  CAS  Google Scholar 

  8. Kroll S, Phillips WJ and Cerione RA (1989) The regulation of the cyclic GMP phosphodiesterase by the GDP-bound form of the α subunit of transducin. J Biol Chem 264: 4490–4497

    PubMed  CAS  Google Scholar 

  9. Lipkin VM, Bondarenko VA, Zagranichny VE, Dobrynina LN, Muradov KhG and Natochin MYu (1993) Site-directed mutagenesis of the cGMP phosphodiesterase γ subunit from bovine rod outer segments: Role of separate amino acid residues in the interaction with catalytic subunits and transducin α subunit. Biochim Biophys Acta 1176:250–256

    Article  PubMed  CAS  Google Scholar 

  10. Lipkin VM, Dumler IL, Muradov KG, Artemyev NO and Etingof RN (1988) Active sites of the cyclic GMP phospho-diesterase γ subunit of retinal rod outer segments. FEBS Lett 234: 287–290

    Article  PubMed  CAS  Google Scholar 

  11. Lipkin VM, Khramtsov NV, Vasilevskaya IA, Atabekova NV, Muradov KG, Gubanov VV, Li T, Johnston JP, Volpp KJ and Applebury ML (1990) The β-subunit of bovine rod photoreceptor cGMP phosphodiesterase: Comparison with the phosphodiesterase family. J Biol Chem 265: 12955–12959

    PubMed  CAS  Google Scholar 

  12. Muradov KhG, Natochin MYu, Bondarenko VA, Skiba NP and Lipkin VM (1990) Interactions between bovine retinal rod cGMP phosphodiesterase subunits. Biol Membr 7: 565–572

    CAS  Google Scholar 

  13. Oppert B, Cunnick JM, Hurt D and Takemoto D (1991) Identification of the retinal cyclic GMP phosphodiesterase inhibitory γ-subunit interaction sites on the catalytic α-subunit. J Biol Chem 266: 16607–16613

    PubMed  CAS  Google Scholar 

  14. Oppert B and Takemoto D (1991) Identification of the γ-subunit interaction sites in the retinal cyclic-GMP phosphodiesterase β-subunit. Biochim Biophys Res Comm 178:474–479

    Article  CAS  Google Scholar 

  15. Otto-Bruc A, Antonny B, Vuong TM, Chardin P and Chabre M (1993) Interaction between the retinal cyclic GMP phosphodiesterase inhibitor and transducin. Kinetics and affinity studies. Biochemistry 32: 8636–8645

    Article  PubMed  CAS  Google Scholar 

  16. Ovchinnikov YuA, Gubanov VV, Khramtsov NV, Ischenko KA, Zagranichny VE, Muradov KG, Shuvaeva TM and Lipkin VM (1987) Cyclic GMP phosphodiesterase from bovine retina. Amino acid sequence of the a subunit and nucleotide sequence of the corresponding cDNA. FEBS Lett 223: 169–173

    Article  PubMed  CAS  Google Scholar 

  17. Ovchinnikov YuA, Lipkin VM, Kumarev VP, Gubanov VV, Khramtsov NV, Akhmedov NB, Zagranichny VE and Muradov KG (1986) Cyclic GMP phosphodiesterase from cattle retina. Amino acid sequence of the β-subunit and nucleotide sequence of the corresponding cDN A. FEBS Lett 204:288–292

    Article  PubMed  CAS  Google Scholar 

  18. Phillips WJ, Trukawinski S and Cerione RA (1989) An antibody-induced enhancement of the transducin-stimulated cyclic GMP phosphodiesterase activity. J Biol Chem 264: 16679–16688

    PubMed  CAS  Google Scholar 

  19. Stryer L (1986) Cyclic GMP cascade in vision. Ann Rev Neurosci 9: 87–119

    Article  PubMed  CAS  Google Scholar 

  20. Takemoto DJ and Cunnick JM (1990) Visual transduction in rod outer segments. Cellular Signalling 2: 99–104

    Article  PubMed  CAS  Google Scholar 

  21. Wensel TG and Stryer L (1986) Reciprocal control of retinal rod cyclic GMP phosphodiesterase by its γ subunit and transducin. PROTEINS: Struct Funct Genet 1: 90–99

    Article  CAS  Google Scholar 

  22. Whalen MM and Bitensky MW (1989) Comparison of the phosphodiesterase inhibitory subunit interactions of frog and bovine rod outer segments. Biochem J 259: 13–19

    PubMed  CAS  Google Scholar 

  23. Whalen MM, Bitensky MW and Takemoto DJ (1990) The effect of the γ-subunit of the cyclic GMP phosphodiesterase of bovine and frog (Rana catesbiana) retinal rod outer segments on the kinetic parameters of the enzyme. Biochem J 265:655–658

    PubMed  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. Branch of Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Pushchino, Moscow Region, 142292, Russia

    V. M. Lipkin, A. M. Alekseev, V. A. Bondarenko, Kh. G. Muradov & V. E. Zagranichny

Authors
  1. V. M. Lipkin
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  2. A. M. Alekseev
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  3. V. A. Bondarenko
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  4. Kh. G. Muradov
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  5. V. E. Zagranichny
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Editor information

Editors and Affiliations

  1. Institute of Biological Sciences, University of Wales, Aberystwyth, UK

    A. R. Smith , A. W. Berry , N. V. J. Harpham  & M. A. Hall , ,  & 

  2. Timiryazev Institute of Plant Physiology, Russian Academy of Sciences, Moscow, Russia

    I. E. Moshkov , G. V. Novikova  & O. N. Kulaeva ,  & 

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© 1996 Kluwer Academic Publishers

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Lipkin, V.M., Alekseev, A.M., Bondarenko, V.A., Muradov, K.G., Zagranichny, V.E. (1996). Site-directed mutagenesis of the cGMP phosphodiesterase inhibitory γ subunit from bovine rods. In: Smith, A.R., et al. Plant Hormone Signal Perception and Transduction. Springer, Dordrecht. https://doi.org/10.1007/978-94-009-0131-5_27

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  • DOI: https://doi.org/10.1007/978-94-009-0131-5_27

  • Publisher Name: Springer, Dordrecht

  • Print ISBN: 978-94-010-6546-7

  • Online ISBN: 978-94-009-0131-5

  • eBook Packages: Springer Book Archive

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