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Methods for Identification of cGKI Substrates

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Guanylate Cyclase and Cyclic GMP

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1020))

Abstract

The cGMP-dependent protein kinases (cGK), which belong to the family of serine/threonine kinases, exhibit their diverse functions in cells through interaction with a variety of substrate proteins. Several substrates were identified and the interactions studied using different methods inter alia co-immunoprecipitation (Co-IP) and cGMP-agarose affinity purification. In the following chapter, we will describe the preparation of cell or tissue lysates, the procedures of cGMP-agarose affinity purification and co-immunoprecipitation, and finally the separation and analysis of the protein complexes by SDS-PAGE or mass spectrometry.

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Acknowledgments

This work was supported by the German Research Council (DFG) and the collaborative research center SFB699.

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Authors and Affiliations

  1. Pharmakologie und Toxikologie, Institut für Pharmazie, Universität Regensburg, Regensburg, Germany

    Katharina Salb & Jens Schlossmann

Authors
  1. Katharina Salb
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  2. Jens Schlossmann
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Editor information

Editors and Affiliations

  1. Addenbrooke’s Hospital, Department of Medicine, University of Cambridge, Hills Road, Cambridge, CB2 2QQ, United Kingdom

    Thomas Krieg

  2. Toxicology and Clinical Pharmacy, Department of Pharmacology, Universität Tübingen, Auf der Morgenstelle 8, Tübingen, 72076, Germany

    Robert Lukowski

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Salb, K., Schlossmann, J. (2013). Methods for Identification of cGKI Substrates. In: Krieg, T., Lukowski, R. (eds) Guanylate Cyclase and Cyclic GMP. Methods in Molecular Biology, vol 1020. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-459-3_9

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  • DOI: https://doi.org/10.1007/978-1-62703-459-3_9

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-458-6

  • Online ISBN: 978-1-62703-459-3

  • eBook Packages: Springer Protocols

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