Skip to main content
SpringerLink
Log in

A Multiparameter Live Cell Imaging Approach to Monitor Cyclic AMP and Protein Kinase A Dynamics in Parallel

  • Protocol
  • First Online:
Fluorescent Protein-Based Biosensors

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1071))

Abstract

Parallel detection of signaling activities allows us to correlate activity dynamics between signaling molecules. In this review, we detail a multiparameter live cell imaging method to monitor 3′,5′-cyclic adenosine monophosphate (cAMP) levels and protein kinase A (PKA) activities in parallel.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Jordan JD, Landau EM, Iyengar R (2000) Signaling networks: the origins of cellular multitasking. Cell 103:193–200

    Article  PubMed  CAS  Google Scholar 

  2. Howe AK (2011) Cross-talk between calcium and protein kinase A in the regulation of cell migration. Curr Opin Cell Biol 23:554–561

    Article  PubMed  CAS  Google Scholar 

  3. Papa S, Bubici C, Zazzeroni F, Franzoso G (2009) Mechanisms of liver disease: cross-talk between the NF-kappaB and JNK pathways. Biol Chem 390:965–976

    Article  PubMed  CAS  Google Scholar 

  4. Kim C, Cheng CY, Saldanha SA, Taylor SS (2007) PKA-I holoenzyme structure reveals a mechanism for cAMP-dependent activation. Cell 130:1032–1043

    Article  PubMed  CAS  Google Scholar 

  5. Seino S, Shibasaki T (2005) PKA-dependent and PKA-independent pathways for cAMP-regulated exocytosis. Physiol Rev 85:1303–1342

    Article  PubMed  CAS  Google Scholar 

  6. Zhou X, Herbst-Robinson KJ, Zhang J (2012) Visualizing dynamic activities of signaling enzymes using genetically encodable FRET-based biosensors from designs to applications. Methods Enzymol 504:317–340

    Article  PubMed  CAS  Google Scholar 

  7. Carlson HJ, Campbell RE (2009) Genetically encoded FRET-based biosensors for multiparameter fluorescence imaging. Curr Opin Biotechnol 20:19–27

    Article  PubMed  CAS  Google Scholar 

  8. Schultz C, Schleifenbaum A, Goedhart J, Gadella TW Jr (2005) Multiparameter imaging for the analysis of intracellular signaling. Chembiochem 6:1323–1330

    Article  PubMed  CAS  Google Scholar 

  9. Shaner NC, Campbell RE, Steinbach PA, Giepmans BN, Palmer AE, Tsien RY (2004) Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nat Biotechnol 22:1567–1572

    Article  PubMed  CAS  Google Scholar 

  10. Aye-Han NN, Allen MD, Ni Q, Zhang J (2012) Parallel tracking of cAMP and PKA signaling dynamics in living cells with FRET-based fluorescent biosensors. Mol Biosyst 8:1435–1440

    Article  PubMed  CAS  Google Scholar 

  11. Allen MD, Zhang J (2008) A tunable FRET circuit for engineering fluorescent biosensors. Angew Chem Int Ed Engl 47:500–502

    Article  PubMed  CAS  Google Scholar 

  12. Allen MD, Zhang J (2006) Subcellular dynamics of protein kinase A activity visualized by FRET-based reporters. Biochem Biophys Res Commun 348:716–721

    Article  PubMed  CAS  Google Scholar 

  13. DiPilato LM, Zhang J (2009) The role of membrane microdomains in shaping beta2-adrenergic receptor-mediated cAMP dynamics. Mol Biosyst 5:832–837

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Nwe-Nwe Aye-Han & Jin Zhang

Authors
  1. Nwe-Nwe Aye-Han
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jin Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Dept. of Pharmacology & Molecular Scienc, Johns Hopkins University, Baltimore, Maryland, USA

    Jin Zhang

  2. Dept. of Pharmacology & Molecular Scienc, Johns Hopkins University, Baltimore, Maryland, USA

    Qiang Ni

  3. Department of Biology, North Caroline A&T State Univ, Greensboro, North Carolina, USA

    Robert H Newman

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Aye-Han, NN., Zhang, J. (2014). A Multiparameter Live Cell Imaging Approach to Monitor Cyclic AMP and Protein Kinase A Dynamics in Parallel. In: Zhang, J., Ni, Q., Newman, R. (eds) Fluorescent Protein-Based Biosensors. Methods in Molecular Biology, vol 1071. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-622-1_16

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-622-1_16

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-621-4

  • Online ISBN: 978-1-62703-622-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation