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Protein Kinase C

Measurement of Translocation, Activation, and Role in Cellular Responses

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Signal Transduction Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 41))

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Abstract

The serine/threonine kinase protein kinase C (PKC) is regarded as playing a key role in stimulation of cellular responses in many different cells and tissues. There are now known to be at least 10 different isoenzymes of PKC (α, βI, βII, γ, δ, ɛ,ζ, η, θ, and λ @#@) which vary in their cofactor requirement for activation and also in their substrate specificity. The conventional PKCs (cPKC) (α, βI, βII, and γ) require Ca2+, phospholipid, and diacylglycerol (DAG) for activation. The novel PKC (nPKC) enzymes (δ, ɛ,ζ, η,and θ) are Ca2+-independent, whereas members of the atypical PKCs (aPKC), ζ and λ, lack the binding site for DAG. Phorbol esters, such as 12-O-tetradecanoylphorbol-13-acetate (TPA), also bind to and activate those enzyme isotypes that are sensitive to DAG and are used as exogenously applied activators of isolated enzymes or of cells to mimic the actions of DAG (1).

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References

  1. Gschwendt, M, Kittstein, W., and Marks, F. (1991) Protein kinase C activation by phorbol esters: do cysteine-rich regions and pseudosubstrate motifs play a role? Trends Biochem. Sci. 16, 167–169.

    Article  PubMed  CAS  Google Scholar 

  2. Nishizuka, Y. (1988) The molecular heterogeneity of protein kinase C and its implications for cellular regulation. Nature 334, 661–665.

    Article  PubMed  CAS  Google Scholar 

  3. Berridge, M. J. and Irvine, R. F. (1984) Inositol trisphosphate, novel second messenger in cellular signal transduction. Nature 312, 315–321.

    Article  PubMed  CAS  Google Scholar 

  4. Wilkinson, S. E. and Hallam, T. J. (1993) Protem kinase C: is its pivotal role in cellular activation over-stated? Trends Pharmacol. Sci. 15(2), 53–57.

    Article  Google Scholar 

  5. Ahmed, S., Kozma, R., Monfries, C., Hall, C., Lim, H. H., Smith, P., and Lim, L. (1990) Human brain n-chimaerm cDNA encodes a novel phorbol ester receptor. Biochem. J. 272, 767–773.

    PubMed  CAS  Google Scholar 

  6. Ono, Y., Fujii, T, Ogita, K., Kikkawa, U., Igarashi, K., and Nishizuka, Y. (1988) The structure, expression and properties of additional members of the protein kinase C family. J. Biol. Chem. 263, 6927–6932.

    PubMed  CAS  Google Scholar 

  7. Liyanage, M, Frith, D., Livneh, E., and Stabel, S. (1992) Protein kinase C group B members PKC-δ, ɛ,ζ, η. Comparison of properties of recombinant proteins in vitro and in vivo. Biochem. J. 283, 781–787.

    PubMed  CAS  Google Scholar 

  8. Borner, C, Nichols-Guadagno, S, Fabbro, D., and Weinstein I. B. (1992) Expression of four protein kinase C isoforms in rat fibroblasts. Differential alterations in ras-, src- and fos-transformed cells. J. Biol. Chem. 267, 12,892–12,899

    PubMed  CAS  Google Scholar 

  9. Nakanishi, H. and Exton, J. H. (1992) Purification and characterization of the zeta-isoform of protein kinase C from bovine kidney. J. Biol. Chem. 267; 16,347–16,354.

    PubMed  CAS  Google Scholar 

  10. Lee, M.-H. and Bell, R. M. (1991) Mechanism of protein kinase C activation by phosphatidylinositol 4,5bisphosphate. Biochem. 30, 1041–1049.

    Article  CAS  Google Scholar 

  11. Ryves, W. J., Evans, A. T., Olivier, A. R., Parker, P. J., and Evans, F. J. (1991) Activation of the PKC-isotypes-α,-βI,-γ,-δ,-ε by phorbol esters of different biological activities. FEBS Lett. 288, 5–9.

    Article  PubMed  CAS  Google Scholar 

  12. Khalil, R. A. and Morgan, K. G. (1991) Imaging of protein kinase C distribution and translocation in living vascular smooth muscle cells Circ. Res. 69, 1626–1631.

    PubMed  CAS  Google Scholar 

  13. Hartwig, J. H., Thelen, M., Rosen, A., Janmey, P. A., Nairn, A. C., and Aderem, A. (1992) MARKS is an actin filament cross-linking protein regulated by protein kinase C and calcium-calmodulin. Nature 356, 618–622.

    Article  PubMed  CAS  Google Scholar 

  14. Isacke, C. M., Meisenhelder, J., Brown, K. D., Gould, K. L., Gould, S. J., and Hunter, T. (1986) Early phosphorylation events following the treatment of Swiss 3T3 cells with bombesin and the mammalian bombesin-related peptide, gastrin-releasing peptide. EMBO J. 5, 2889–2898.

    PubMed  CAS  Google Scholar 

  15. Tyers, M., Rachubinski, R. A., Sterwart, M. I., Varrichio, A. M., Shorr, R. G., Haslam, R. J., and Harley, C. B. (1988) Molecular cloning and expression of the major protein kinase C substrate of platelets. Nature 333, 470–473.

    Article  PubMed  CAS  Google Scholar 

  16. Bradshaw, D., Hill, C. H., Nixon, J. S., and Wilkinson, S. E. (1993) Therapeutic potential of protein kinase C inhibitors. Agents and Actions, 38, 135–147.

    Article  PubMed  CAS  Google Scholar 

  17. Nixon, J. S., Wilkinson, S. E., Davis, P. D., Sedgwick, A. D., Wadsworth, J., and Westmacott, D. (1991) Modulation of cellular processes by H7, a non-selective inhibitor of protein kinases. Agents and Actions 32, 188–192.

    Article  PubMed  CAS  Google Scholar 

  18. Birchall, A. M., Bishop, J., Bradshaw, D., Cline A., Coffey, J., Elliott, L. H., et al. (1994) Ro 32-0432, a selective and orally active inhibitor of protein kinase C prevents T-cell activation. J. Pharm. Exp. Ther. 268(2), 922–929.

    CAS  Google Scholar 

  19. Toullec, D., Pianetti, P., Coste, H., Bellevergue, P., Grand, T., Ajakane, M., et al. (1991) The bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase C. J. Biol. Chem. 266, 15,771–15,781.

    PubMed  CAS  Google Scholar 

  20. Davis, P. D., Hill, C. H., Keech, E., Lawton, G., Nixon, J. S., Sedgwick, A. D., et al. (1989) Potent selective inhibitors of protein kinase C. FEBS Lett. 259, 61–63.

    Article  PubMed  CAS  Google Scholar 

  21. Hug, H. and Sarre, T. F. (1993) Protein kinase C isoenzymes: divergence in signal transduction? Biochem. J. 291, 329–343

    PubMed  CAS  Google Scholar 

  22. Wilkinson, S. E., Parker, P. J., and Nixon, J. S. (1993) Isoenzyme specificity of bisindolylmaleimide, selective inhibitors of protein kinase C. Biochem. J. 294, 335–337

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Roche Research Centre, Hertfordshire, UK

    Sandra E. Wilkinson & Trevor J. Hallam

Authors
  1. Sandra E. Wilkinson
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  2. Trevor J. Hallam
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Editor information

Editors and Affiliations

  1. Queen’s Medical Centre, University of Nottingham, UK

    David A. Kendall  & Stephen J. Hill  & 

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© 1995 Humana Press Inc , Totowa, NJ

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Wilkinson, S.E., Hallam, T.J. (1995). Protein Kinase C. In: Kendall, D.A., Hill, S.J. (eds) Signal Transduction Protocols. Methods in Molecular Biology™, vol 41. Humana Press. https://doi.org/10.1385/0-89603-298-1:261

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  • DOI: https://doi.org/10.1385/0-89603-298-1:261

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-298-9

  • Online ISBN: 978-1-59259-528-0

  • eBook Packages: Springer Protocols

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