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A Live-Cell Imaging Assay for Nuclear Entry of cAMP-Dependent Protein Kinase Catalytic Subunits Stimulated by Endogenous GPCR Activation

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cAMP Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2483))

Abstract

Nuclear entry of cAMP-dependent protein kinase catalytic subunits is typically inferred from changes in net protein amount or kinase activity in the nucleus. Previous methods to directly assess nuclear entry require kinase subunit overexpression and/or supraphysiological cAMP elevation. We describe a method to detect nuclear entry of catalytic subunits expressed at an endogenous level in living cells, stimulated by cAMP in a physiological range, and in real time.

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Author information

Authors and Affiliations

  1. Program in Cell Biology, University of California, San Francisco, San Francisco, CA, USA

    Grace E. Peng

  2. Department of Psychiatry and Behavioral Sciences, University of California, San Francisco, San Francisco, CA, USA

    Grace E. Peng & Mark von Zastrow

  3. Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA, USA

    Mark von Zastrow

  4. Quantitative Biology Institute, University of California, San Francisco, San Francisco, CA, USA

    Mark von Zastrow

Authors
  1. Grace E. Peng
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  2. Mark von Zastrow
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Corresponding author

Correspondence to Mark von Zastrow .

Editor information

Editors and Affiliations

  1. Department of Physiology, Anatomy and Genetics, Oxford University, Oxford, UK

    Manuela Zaccolo

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Peng, G.E., von Zastrow, M. (2022). A Live-Cell Imaging Assay for Nuclear Entry of cAMP-Dependent Protein Kinase Catalytic Subunits Stimulated by Endogenous GPCR Activation. In: Zaccolo, M. (eds) cAMP Signaling. Methods in Molecular Biology, vol 2483. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2245-2_21

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  • DOI: https://doi.org/10.1007/978-1-0716-2245-2_21

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2244-5

  • Online ISBN: 978-1-0716-2245-2

  • eBook Packages: Springer Protocols

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