Role of membrane microdomains in compartmentation of cAMP signaling

Shailesh R. Agarwal, Pei Chi Yang, Monica Rice, Cherie A. Singer, Viacheslav O. Nikolaev, Martin J. Lohse, Colleen E Clancy, Robert D. Harvey

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Spatially restricting cAMP production to discrete subcellular locations permits selective regulation of specific functional responses. But exactly where and how cAMP signaling is confined is not fully understood. Different receptors and adenylyl cyclase isoforms responsible for cAMP production are not uniformly distributed between lipid raft and non-lipid raft domains of the plasma membrane. We sought to determine the role that these membrane domains play in organizing cAMP responses in HEK293 cells. The freely diffusible FRET-based biosensor Epac2-camps was used to measure global cAMP responses, while versions of the probe targeted to lipid raft (Epac2-MyrPalm) and non-raft (Epac2-CAAX) domains were used to monitor local cAMP production near the plasma membrane. Disruption of lipid rafts by cholesterol depletion selectively altered cAMP responses produced by raft-associated receptors. The results indicate that receptors associated with lipid raft as well as non-lipid raft domains can contribute to global cAMP responses. In addition, basal cAMP activity was found to be significantly higher in non-raft domains. This was supported by the fact that pharmacologic inhibition of adenylyl cyclase activity reduced basal cAMP activity detected by Epac2-CAAX but not Epac2-MyrPalm or Epac2-camps. Responses detected by Epac2-CAAX were also more sensitive to direct stimulation of adenylyl cyclase activity, but less sensitive to inhibition of phosphodiesterase activity. Quantitative modeling was used to demonstrate that differences in adenylyl cyclase and phosphodiesterase activities are necessary but not sufficient to explain compartmentation of cAMP associated with different microdomains of the plasma membrane.

Original languageEnglish (US)
Article numbere95835
JournalPLoS One
Volume9
Issue number4
DOIs
StatePublished - Apr 21 2014

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Membrane Microdomains
adenylate cyclase
Adenylyl Cyclases
Cell membranes
Membranes
Lipids
plasma membrane
Phosphoric Diester Hydrolases
Cell Membrane
lipids
receptors
biosensors
HEK293 Cells
Biosensing Techniques
Biosensors
Protein Isoforms
Cholesterol
cholesterol
monitoring
cells

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Agarwal, S. R., Yang, P. C., Rice, M., Singer, C. A., Nikolaev, V. O., Lohse, M. J., ... Harvey, R. D. (2014). Role of membrane microdomains in compartmentation of cAMP signaling. PLoS One, 9(4), [e95835]. https://doi.org/10.1371/journal.pone.0095835

Role of membrane microdomains in compartmentation of cAMP signaling. / Agarwal, Shailesh R.; Yang, Pei Chi; Rice, Monica; Singer, Cherie A.; Nikolaev, Viacheslav O.; Lohse, Martin J.; Clancy, Colleen E; Harvey, Robert D.

In: PLoS One, Vol. 9, No. 4, e95835, 21.04.2014.

Research output: Contribution to journalArticle

Agarwal, SR, Yang, PC, Rice, M, Singer, CA, Nikolaev, VO, Lohse, MJ, Clancy, CE & Harvey, RD 2014, 'Role of membrane microdomains in compartmentation of cAMP signaling', PLoS One, vol. 9, no. 4, e95835. https://doi.org/10.1371/journal.pone.0095835
Agarwal SR, Yang PC, Rice M, Singer CA, Nikolaev VO, Lohse MJ et al. Role of membrane microdomains in compartmentation of cAMP signaling. PLoS One. 2014 Apr 21;9(4). e95835. https://doi.org/10.1371/journal.pone.0095835
Agarwal, Shailesh R. ; Yang, Pei Chi ; Rice, Monica ; Singer, Cherie A. ; Nikolaev, Viacheslav O. ; Lohse, Martin J. ; Clancy, Colleen E ; Harvey, Robert D. / Role of membrane microdomains in compartmentation of cAMP signaling. In: PLoS One. 2014 ; Vol. 9, No. 4.
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