Live-cell Measurement of Odorant Receptor Activation Using a Real-time cAMP Assay.

Journal Article (Journal Article)

The enormous sizes of the mammalian odorant receptor (OR) families present difficulties to find their cognate ligands among numerous volatile chemicals. To efficiently and accurately deorphanize ORs, we combine the use of a heterologous cell line to express mammalian ORs and a genetically modified biosensor plasmid to measure cAMP production downstream of OR activation in real time. This assay can be used to screen odorants against ORs and vice versa. Positive odorant-receptor interactions from the screens can be subsequently confirmed by testing against various odor concentrations, generating concentration-response curves. Here we used this method to perform a high-throughput screening of an odorous compound against a human OR library expressed in Hana3A cells and confirmed that the positively-responding receptor is the cognate receptor for the compound of interest. We found this high-throughput detection method to be efficient and reliable in assessing OR activation and our data provide an example of its potential use in OR functional studies.

Full Text

Duke Authors

Cited Authors

  • Zhang, Y; Pan, Y; Matsunami, H; Zhuang, H

Published Date

  • October 2, 2017

Published In

PubMed ID

  • 28994818

Pubmed Central ID

  • PMC5752358

Electronic International Standard Serial Number (EISSN)

  • 1940-087X

Digital Object Identifier (DOI)

  • 10.3791/55831

Language

  • eng

Conference Location

  • United States