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

Publication ,  Journal Article
Fukutani, Y; Ishii, J; Kondo, A; Ozawa, T; Matsunami, H; Yohda, M
Published in: Biotechnol Bioeng
June 2017

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.

Duke Scholars

Published In

Biotechnol Bioeng

DOI

EISSN

1097-0290

Publication Date

June 2017

Volume

114

Issue

6

Start / End Page

1354 / 1361

Location

United States

Related Subject Headings

  • Saccharomyces cerevisiae
  • Receptors, Somatostatin
  • Receptors, G-Protein-Coupled
  • Luminescent Measurements
  • Luciferases
  • Genes, Reporter
  • Biotechnology
  • Biological Assay
 

Citation

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Fukutani, Y., Ishii, J., Kondo, A., Ozawa, T., Matsunami, H., & Yohda, M. (2017). Split luciferase complementation assay for the analysis of G protein-coupled receptor ligand response in Saccharomyces cerevisiae. Biotechnol Bioeng, 114(6), 1354–1361. https://doi.org/10.1002/bit.26255
Fukutani, Yosuke, Jun Ishii, Akihiko Kondo, Takeaki Ozawa, Hiroaki Matsunami, and Masafumi Yohda. “Split luciferase complementation assay for the analysis of G protein-coupled receptor ligand response in Saccharomyces cerevisiae.Biotechnol Bioeng 114, no. 6 (June 2017): 1354–61. https://doi.org/10.1002/bit.26255.
Fukutani Y, Ishii J, Kondo A, Ozawa T, Matsunami H, Yohda M. Split luciferase complementation assay for the analysis of G protein-coupled receptor ligand response in Saccharomyces cerevisiae. Biotechnol Bioeng. 2017 Jun;114(6):1354–61.
Fukutani, Yosuke, et al. “Split luciferase complementation assay for the analysis of G protein-coupled receptor ligand response in Saccharomyces cerevisiae.Biotechnol Bioeng, vol. 114, no. 6, June 2017, pp. 1354–61. Pubmed, doi:10.1002/bit.26255.
Fukutani Y, Ishii J, Kondo A, Ozawa T, Matsunami H, Yohda M. Split luciferase complementation assay for the analysis of G protein-coupled receptor ligand response in Saccharomyces cerevisiae. Biotechnol Bioeng. 2017 Jun;114(6):1354–1361.
Journal cover image

Published In

Biotechnol Bioeng

DOI

EISSN

1097-0290

Publication Date

June 2017

Volume

114

Issue

6

Start / End Page

1354 / 1361

Location

United States

Related Subject Headings

  • Saccharomyces cerevisiae
  • Receptors, Somatostatin
  • Receptors, G-Protein-Coupled
  • Luminescent Measurements
  • Luciferases
  • Genes, Reporter
  • Biotechnology
  • Biological Assay