title: Split luciferase complementation assay for the analysis of G protein-coupled receptor ligand response in Saccharomyces cerevisiae.
abstract: 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.
authors.affiliation: Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Koganei, Tokyo, 184-8588, Japan.
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Fukutani
authors.affiliation: Graduate School of Science, Technology and Innovation, Kobe university, Kobe, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Ishii
authors.affiliation: Graduate School of Science, Technology and Innovation, Kobe university, Kobe, Japan.
authors.forename: Akihiko
authors.identifier: 0000-0003-1527-5288
authors.initials: A
authors.lastname: Kondo
authors.affiliation: Department of Chemistry, School of Science, The University of Tokyo, Hongo, Tokyo, Japan.
authors.forename: Takeaki
authors.initials: T
authors.lastname: Ozawa
authors.affiliation: Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina.|Institute of Global Innovation Research, Tokyo University of Agriculture and Technology, Koganei, Tokyo, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Matsunami
authors.affiliation: Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Koganei, Tokyo, 184-8588, Japan.|Institute of Global Innovation Research, Tokyo University of Agriculture and Technology, Koganei, Tokyo, Japan.
authors.forename: Masafumi
authors.identifier: 0000-0001-8307-9671
authors.initials: M
authors.lastname: Yohda
mesh_terms: D001681:Biological Assay / Q000379:methods*; D017930:Genes, Reporter / Q000235:genetics; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D008163:Luminescent Measurements / Q000379:methods*; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*; D017481:Receptors, Somatostatin / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism
chemical_list: D043562:Receptors, G-Protein-Coupled; D017481:Receptors, Somatostatin; C574734:SSTR2 protein, human; D008156:Luciferases
keywords: G protein coupled receptor; biosensing; split-luciferase