Split luciferase complementation assay for the analysis of G protein-coupled receptor ligand response in Saccharomyces cerevisiae.

Published

Journal Article

The budding yeast Saccharomyces cerevisiae is equipped with G protein-coupled receptors (GPCR). Because the yeast GPCR signaling mechanism is partly similar to that of the mammalian system, S. cerevisiae can be used for a host of mammalian GPCR expression and ligand-mediated activation assays. However, currently available yeast systems require several hours to observe the responses because they depend on the expression of reporter genes. In this study, we attempted to develop a simple GPCR assay system using split luciferase and β-arrestin, which are independent of the endogenous S. cerevisiae GPCR signaling pathways. We applied the split luciferase complementation assay method to S. cerevisiae and found that it can be used to analyze the ligand response of the human somatostatin receptor in S. cerevisiae. On the contrary, the response of the pheromone receptor Ste2 was not observed by the assay. Thus, the split luciferase complementation should be free from the effect of the endogenous GPCR signaling. Biotechnol. Bioeng. 2017;114: 1354-1361. © 2017 Wiley Periodicals, Inc.

Full Text

Duke Authors

Cited Authors

  • Fukutani, Y; Ishii, J; Kondo, A; Ozawa, T; Matsunami, H; Yohda, M

Published Date

  • June 2017

Published In

Volume / Issue

  • 114 / 6

Start / End Page

  • 1354 - 1361

PubMed ID

  • 28112390

Pubmed Central ID

  • 28112390

Electronic International Standard Serial Number (EISSN)

  • 1097-0290

Digital Object Identifier (DOI)

  • 10.1002/bit.26255

Language

  • eng

Conference Location

  • United States