148
Views
17
CrossRef citations to date
0
Altmetric
Research Article

Label-free optical biosensor for probing integrative role of adenylyl cyclase in G protein-coupled receptor signaling

&
Pages 154-162 | Received 22 Dec 2008, Accepted 17 May 2009, Published online: 16 Jul 2009
 

Abstract

Adenylyl cyclase is considered as an integrator for receptor signaling. However, its integrative role in receptor signaling is largely studied at the level of point of contacts in complex pathways. Here we used forskolin as a pharmacological probe and the resonant waveguide grating (RWG) biosensor to examine the signal integration of G protein-coupled receptors (GPCRs) at the cyclase-cyclic AMP-PKA module. The biosensor is a refractive index sensitive optical biosensor that is capable of detecting ligand-induced dynamic mass redistribution in cells without labels and cellular manipulations. Stimulation of seven cell lines with forskolin led to distinct optical responses, indicative of distinct expressions and/or organization of cyclase isoforms. The forskolin response in A431 was sensitive to the activities of protein kinase A, Rho kinase, and MAP kinases. Desensitization assays showed that the forskolin pretreatment heterologously desensitized Gs signaling, partially attenuated Gq signaling, but had complicate impacts on Gi signaling. This study documents the integrative role of adenylyl cyclase in GPCR signaling and the power of forskolin as a pharmacological probe to differentiate receptor signaling using the label-free biosensor cellular assays.

Acknowledgements

Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.

Log in via your institution

Log in to Taylor & Francis Online

PDF download + Online access

  • 48 hours access to article PDF & online version
  • Article PDF can be downloaded
  • Article PDF can be printed
USD 65.00 Add to cart

Issue Purchase

  • 30 days online access to complete issue
  • Article PDFs can be downloaded
  • Article PDFs can be printed
USD 1,339.00 Add to cart

* Local tax will be added as applicable

Related Research

People also read lists articles that other readers of this article have read.

Recommended articles lists articles that we recommend and is powered by our AI driven recommendation engine.

Cited by lists all citing articles based on Crossref citations.
Articles with the Crossref icon will open in a new tab.