Development of Highly Specific, Membrane Permeable Peptide Blockers of cGMP-Dependent Protein Kinase Utilizing Affinity Selection from Peptide Libraries

  • Werner Tegge
  • Ronald Frank
  • Mark S. Taylor
  • Joseph E. Brayden
  • Christian K. Nickl
  • Wolfgang R. G. Dostmann
Part of the American Peptide Symposia book series (APSY, volume 7)

Abstract

The cGMP-dependent protein kinases type Iα and Iβ (cGPK) act directly downstream in the nitric oxide (NO) mediated signaling pathway in the control of a variety of cellular responses, ranging from smooth muscle cell relaxation to neuronal synaptic plasticity [1,2]. The structural similarity of cGPK and its closest relative, the cAMP-dependent protein kinase (cAPK), has made it difficult to study cGPK pathways independent of those mediated by cAPK, primarily due to the lack of potent and selective cGPK inhibitors. Here we report a novel peptide library screen specifically designed to select for tight binding peptides that identified selective inhibitors of the kinases. The most potent sequences were made cell permeable by the addition of membrane translocation sequences from HIV and Drosophila and physiological effects of the constructs were studied in vivo and in vitro.

Keywords

Cytosol Fluorescein 

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References

  1. 1.
    Lincoln, T.M., Komalavilas, P., Boerth, N.J., Mac-Millan-Crow, L.A., Cornwell, T.L. Adv. Pharmacol 34, 305–322 (1995).PubMedCrossRefGoogle Scholar
  2. 2.
    Pfeifer, A., Ruth, P., Dostmann, W., Sausbier, M., Klatt, P., Hofmann, F. Rev. Physiol. Biochem. Pharmacol. 135, 105–149 (1999).PubMedCrossRefGoogle Scholar
  3. 3.
    Part of this work has been published elsewhere: Dostmann, W.R.G., Taylor, M.S., Nickl, C.K., Brayden, J.E., Frank, R., Tegge, W. Proc. Natl. Acad. Sci. U.S.A. 97, 14772–14777 (2000).PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2001

Authors and Affiliations

  • Werner Tegge
    • 1
  • Ronald Frank
    • 1
  • Mark S. Taylor
    • 2
  • Joseph E. Brayden
    • 2
  • Christian K. Nickl
    • 2
  • Wolfgang R. G. Dostmann
    • 2
  1. 1.German Research Centre for Biotechnology (GBF)AG Molecular RecognitionBraunschweigGermany
  2. 2.Departments of Pharmacology and Molecular PhysiologyUniversity of Vermont, College of MedicineBurlingtonUSA

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