Protein Phosphatase Protocols pp 41-58

Part of the Methods in Molecular Biology™ book series (MIMB, volume 93)

Inhibitor-1, a Regulator of Protein Phosphatase 1 Function

  • John H. Connor
  • Hai Quan
  • Carey Oliver
  • Shirish Shenolikar
Protocol

Abstract

With the discovery of numerous eukaryotic protein serine/threonine phosphatases, a classification scheme was developed that exploited some of the biochemical properties of these enzymes to expedite their identification (1). These properties included the preferential dephosphorylation of either α or β-subunits of phosphorylase kinase, the metal ion dependency displayed by some phosphatases for enzyme activity and their inhibition by two mammalian thermostable proteins, inhibitor-1 (I-1) and inhibitor-2 (I-2).

References

  1. 1.
    Ingebritsen, T. S. and Cohen, P. (1983) The protein phosphatases involved in cellular regulation. 1. Classification and substrate specificities. Eur. J. Biochem. 132, 244–261Google Scholar
  2. 2.
    Stewart, A.A, Ingebritsen, T S, and Cohen, P (1983) The protein phosphatases involved in cellular regulation 5. Purification and properties of a Ca2+/calmodulin-dependent protein phosphatase (2B) from rabbit skeletal muscle Eur J Biochem 132, 289–295PubMedCrossRefGoogle Scholar
  3. 3.
    Klee, C B., Krinks, M H., Manalan, A S., Cohen, P, and Stewart, A A. (1983) Isolation and characterization of bovine brain calcineurin: a calmodulin-stimulated protein phosphatase Methods Enzymol 102, 227–244PubMedCrossRefGoogle Scholar
  4. 4.
    Alessi, D, Street, A J., Cohen, P, and Cohen, P T W (1993) Inhibitor-2 functions like a chaperone to fold three expressed isoforms of mammalian protein phosphatase-1 into a conformation with the specificity and regulatory properties of the native enzyme Eur J Biochem 213, 1055–1066PubMedCrossRefGoogle Scholar
  5. 5.
    Brautigan, D L, Gruppuso, P. A., and Mumby, M (1986) Protein phosphatase type-1 and type-2 catalytic subunits both bind inhibitor-2 and monoclonal antibodies J Biol Chem 261, 14,924–14,928PubMedGoogle Scholar
  6. 6.
    Mulkey, R M, Endo, S., Shenolikar, S, and Malenka, R. C (1994) Involvement of a calcineurin/inhibitor-1 phosphatase cascade in hippocampal long-term depression Nature 369, 486–488PubMedCrossRefGoogle Scholar
  7. 7.
    Endo, S., Critz, S, Byrne, J. H., and Shenolikar, S. (1995) Protein phosphatase-1 regulates outward K+ currents in sensory neurons in Aplysia californica J Neurochem 64, 1833–1840.CrossRefGoogle Scholar
  8. 8.
    Foulkes, J. G. and Cohen, P. (1981) The hormonal control of glycogen metabolism Phosphorylation of protein phosphatase inhibitor-1 in vivo in response to adrenaline. Eur J. Biochem 97, 251–256CrossRefGoogle Scholar
  9. 9.
    Foulkes, J G., Cohen, P., Strada, S. J., Everson, W V, and Jefferson, L S. (1982) Antagonistic effects of insulin and beta-adrenergic agonists on the activity of protein phosphatase inhibitor-1 in skeletal muscle of the perfused rat hemicorpus J Biol Chem 257, 12,493–12,496PubMedGoogle Scholar
  10. 10.
    Khatra, B. S, Chiasson, J. L, Shikama, H, Exton, J H., and Soderling, T R (1980) Effect of epinephrine and insulin on the dephosphorylation of protein phosphatase inhibitor-1 in perfused rat skeletal muscle. FEBS Lett. 114, 253–256PubMedCrossRefGoogle Scholar
  11. 11.
    Ahmad, Z, Green, F J., Subuhi, H S, and Watanabe, A M (1989) Autonomic regulation of type 1 protein phosphatase in cardiac muscle. J Biol Chem 264, 3859–3866.PubMedGoogle Scholar
  12. 12.
    Neumann, J, Gupta, R. C, Schmitz, W., Scholz, H, Nairn A. C, and Watanabe, A. M. (1991) Evidence for isoproterenol-induced phosphorylation of phosphatase inhibitor-1 in intact heart Circulation Res. 69, 1450–1457PubMedGoogle Scholar
  13. 13.
    Nemenoff, R. A, Blackshear, P J., and Avruch, J (1983) Hormonal regulation of protein dephosphorylation. Identification and hormonal regulation of protein phosphatase inhibitor-1 in rat adipose tissue. J. Biol Chem. 258, 9437–9443PubMedGoogle Scholar
  14. 14.
    Snyder, G. L., Girault, J. A, Chen, J. Y., Czernik, A. J., Kebabian, J. W., Nathanson, J A, and Greengard, P (1992) Phosphorylation of DARPP-32 and protein phosphatase inhibitor-1 in rat choroid plexus: regulation by factors other than dopamine J Neurosci 12, 3071–3083.PubMedGoogle Scholar
  15. 15.
    MacDougall, L K., Campbell, D. G., Hubbard, M. J., and Cohen, P. (1989) Partial structure and hormonal regulation of rabbit liver inhibitor-1; distribution of inhibitor-1 in rabbit and rat tissues Biochim Biophys. Acta 1010, 218–226.PubMedCrossRefGoogle Scholar
  16. 16.
    Elbrecht, A, DiRenzo, J., Smith, R G, and Shenolikar, S (1990) Molecular cloning of the cDNA for protein phosphatase inhibitor-1 and expression in rat and rabbit tissues J Biol Chem 265, 13,415–13,418PubMedGoogle Scholar
  17. 17.
    Endo, S, Zhou, X. Z., Connor, J., Wang, B., and Shenolikar, S (1996) Multiple structural elements define the specificity of recombinant human inhibitor-1 as a protein phosphatase-1 inhibitor. Biochemistry 35, 5220–5228.PubMedCrossRefGoogle Scholar
  18. 18.
    Nimmo, G A and Cohen, P (1978) The regulation of glycogen metabolism Purification and characterization of protein phosphatase inhibitor-1 from rabbit skeletal muscle Eur J Biochem 87, 341–351PubMedCrossRefGoogle Scholar
  19. 19.
    Shenolikar, S. and Ingebritsen, T S (1984) Protein (senne,threonine) phosphatases. Methods Enzymol 107, 102–129.PubMedCrossRefGoogle Scholar
  20. 20.
    Cohen, P, Foulkes, J. G, Holmes, C. F, Nimmo, G A, and Tonks, N K (1988) Protein phosphatase inhibitor-1 and inhibitor-2 from rabbit skeletal muscle. Methods Enzymol 159, 427–437.PubMedCrossRefGoogle Scholar
  21. 21.
    Tartof, K. D and Hobbs, C. A (1988) New cloning vectors and techniques for easy and rapid restriction mapping Gene 67, 169–182PubMedCrossRefGoogle Scholar
  22. 22.
    Tabor, S and Richardson, C C (1992) A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes Biotechnology 24, 280–284.PubMedGoogle Scholar
  23. 23.
    Foulkes, J. G, Strada, S J., Henderson, P J, and Cohen, P (1983) A kinetic analysis of the effects of inhibitor-1 and inhibitor-2 on the activity of protein phosphatase-1. Eur J Biochem 132, 309–313.PubMedCrossRefGoogle Scholar
  24. 24.
    Beavo, J A, Bechtel, P. J, and Krebs, E G (1974) Preparation of homogenous cyclic AMP-dependent protein kinase(s) and its subunits from rabbit skeletal muscle Methods Enzymol 38, 299–308PubMedCrossRefGoogle Scholar
  25. 25.
    Cohen, P. (1989) The structure and regulation of protein phosphatases Annu Rev Biochem 58, 453–508PubMedCrossRefGoogle Scholar
  26. 26.
    Shenolikar, S. and Nairn, A. (1991) Protein phosphatases recent progress Adv Second Messenger Phosphoprotein Res 23, 1–121PubMedGoogle Scholar
  27. 27.
    Bollen, M and Stalmans, W. (1992) The structure, role and regulation of type-1 protein phosphatases. Crit. Rev. Biochem Mol Biol. 27, 227–281PubMedCrossRefGoogle Scholar
  28. 28.
    Shenolikar, S. (1994) Protein serine/threonine phosphatases, new avenues for cell regulation Annu Rev Cell Biol 10, 55–86.PubMedCrossRefGoogle Scholar
  29. 29.
    Gratecos, D. and Fischer, E. H (1974) Adenosine 5′-o(3-thiotriphosphate) in the control of phosphorylase activity. Biochem Biophys. Res. Commun 58, 960–967.PubMedCrossRefGoogle Scholar
  30. 30.
    McGowen, C. H. and Cohen, P (1988) Protein phosphatase-2C from rabbit skeletal muscle and liver: an Mg2+-dependent enzyme. Methods Enzymol. 159, 416–426CrossRefGoogle Scholar
  31. 31.
    Pato, M. D. and Kerc, E (1988) Purification of smooth muscle myosin phosphatase from turkey gizzard Methods Enzymol. 159, 446–453.PubMedCrossRefGoogle Scholar
  32. 3.
    Tonks, N. K., Diltz, C D., and Fischer, E H (1991) Purification of protein-tyrosine phosphatases from human placenta. Methods Enzymol 201, 427–442.PubMedCrossRefGoogle Scholar
  33. 33.
    Yang, S. D., Vandenheede, J R., and Merlevede, W. (1981) A simplified procedure for the purification of the protein phosphatase modulator (inhibitor-2) from rabbit skeletal muscle. FEBS Lett. 132, 293–295PubMedCrossRefGoogle Scholar
  34. 34.
    Holmes, C F B and Boland, M. (1993) Inhibitors of protein phosphatase-1 and-2A; two major serine-threonine phosphatases involved in cellular regulation. Curr Biol 3, 934–943CrossRefGoogle Scholar
  35. 35.
    Cohen, P., Nimmo, G A., Shenolikar, S., and Foulkes, J. G. (1979) The role of inhibitor-1 in the cyclic AMP mediated control of glycogen metabolism in skeletal muscle. “Cyclic Nucleotides and Protein Phosphorylation in Cell Regulation” (Krause, E.G., ed) Pergamon, Oxford, FEBS Symp 54, 161–169Google Scholar
  36. 36.
    Shenolikar, S., Foulkes J G, and Cohen, P (1978) Isolation of an active fragment of protein phosphatase inhibitor-1 from rabbit skeletal muscle Biochem Soc. Trans. 6, 111–113.Google Scholar
  37. 37.
    Aitken, A and Cohen, P. (1984) Isolation and characterization of active fragments of protein phosphatase inhibitor-1 from rabbit skeletal muscle. FEBS Lett. 147, 54–58.CrossRefGoogle Scholar
  38. 38.
    Hemmings, H C. Jr., Nairn, A C, Elliott, J. I., and Greengard, P. (1990) Synthetic peptide analogs of DARPP-32 (Mr 32,000 dopamine-and cAMP-regulated phosphoprotein), an inhibitor of protein phosphatase-1 J. Biol Chem 265, 20,369–20,376PubMedGoogle Scholar
  39. 39.
    Cohen, P (1988) Protein phosphorylation and hormone action. Proc Royal Soc Lond Biol. Sci. 234, 115–144CrossRefGoogle Scholar
  40. 40.
    Nakielny, S, Campbell, D. G, and Cohen, P. (1991) The molecular mechanism by which adrenalin inhibits glycogen synthesis. Eur J Biochem. 199, 713–722PubMedCrossRefGoogle Scholar
  41. 41.
    Gupta, R. C, Neumann, J., and Watanabe, A. M (1993) Comparison of adenosine and muscarinic receptor-mediated effects on protein phosphatase inhibitor-1 activity in the heart. J Pharmacol. Exp Ther 266, 16–22.PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 1998

Authors and Affiliations

  • John H. Connor
    • 1
  • Hai Quan
    • 1
  • Carey Oliver
    • 1
  • Shirish Shenolikar
    • 1
  1. 1.Department of Pharmacology and Cancer BiologyDuke University Medical CenterDurham

Personalised recommendations