| Title: | Successful expression of a functional yeast G-protein-coupled receptor (Ste2) in mammalian cells |
| Author(s): | Yin D; Gavi S; Shumay E; Duell K; Konopka JB; Malbon CC; Wang HY; |
| Address: | "Department of Molecular Pharmacology, University Medical Center, SUNY/Stony Brook, Stony Brook, NY 11794-8651, USA" |
| Journal Title: | Biochem Biophys Res Commun |
| DOI: | 10.1016/j.bbrc.2005.01.130 |
| ISSN/ISBN: | 0006-291X (Print) 0006-291X (Linking) |
| Abstract: | "G-protein-coupled receptors (GPCRs) are membrane-embedded cell signaling devices transducing ligand binding to activation of heterotrimeric G-proteins, providing a paradigm for signaling for yeast and mammals alike. Probing the extent to which yeast GPCRs may couple to mammalian G-proteins has been problematic. In the current work, we explored conditions that enable the cell-surface expression of a yeast alpha-factor pheromone receptor (Ste2). When expressed in human HEK293 cells, Ste2 is shown to bind its ligand alpha-factor, to be functional and catalyze activation of the mitogen-activated protein kinase cascade, and to demonstrate agonist-induced internalization. In response to agonist Ste2 as maintained intracellularly for several hours and avoids the degradation process observed for Ste2 in yeast cells. This is the first successful demonstration of the ability to express a functional yeast GPCR in mammalian cells" |
| Keywords: | "Animals Biochemistry/*methods CHO Cells Catalysis Cell Line Cricetinae Green Fluorescent Proteins/metabolism Humans Immunoblotting Ligands MAP Kinase Signaling System Microscopy, Confocal Mitogen-Activated Protein Kinase 1/metabolism Mitogen-Activated Pro;" |
| Notes: | "MedlineYin, Dezhong Gavi, Shai Shumay, Elena Duell, Ken Konopka, James B Malbon, Craig C Wang, Hsien-Yu eng Research Support, U.S. Gov't, P.H.S. 2005/02/22 Biochem Biophys Res Commun. 2005 Apr 1; 329(1):281-7. doi: 10.1016/j.bbrc.2005.01.130" |
