« Previous Abstract"Synthesis of the Enantiomers of (Z)-5-(1-Octenyl)oxacyclopentan-2-one, a Sex Pheromone of the Cupreous Chafer Beetle, Anomala cuprea Hope"    Next AbstractVolatile profile and microbiological characterization of hollow defect in dry-cured ham »

Biotechnol Bioeng

Title:		Split luciferase complementation assay for the analysis of G protein-coupled receptor ligand response in Saccharomyces cerevisiae
Author(s):		Fukutani Y; Ishii J; Kondo A; Ozawa T; Matsunami H; Yohda M;
Address:		"Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Koganei, Tokyo, 184-8588, Japan. Graduate School of Science, Technology and Innovation, Kobe university, Kobe, Japan. Department of Chemistry, School of Science, The University of Tokyo, Hongo, Tokyo, Japan. 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"
Journal Title:		Biotechnol Bioeng
Year:		2017
Volume:		20170314
Issue:		6
Page Number:		1354 - 1361
DOI:		10.1002/bit.26255
ISSN/ISBN:		1097-0290 (Electronic) 0006-3592 (Linking)
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 beta-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. (c) 2017 Wiley Periodicals, Inc"
Keywords:		"Biological Assay/*methods Genes, Reporter/genetics Luciferases/genetics/*metabolism Luminescent Measurements/*methods Receptors, G-Protein-Coupled/genetics/*metabolism Receptors, Somatostatin/genetics/*metabolism Saccharomyces cerevisiae/genetics/metaboli;"
Notes:		"MedlineFukutani, Yosuke Ishii, Jun Kondo, Akihiko Ozawa, Takeaki Matsunami, Hiroaki Yohda, Masafumi eng 2017/01/24 Biotechnol Bioeng. 2017 Jun; 114(6):1354-1361. doi: 10.1002/bit.26255. Epub 2017 Mar 14"