-
Views
-
Cite
Cite
Keisuke Hara, Takuya Ono, Kouichi Kuroda, Mitsuyoshi Ueda, Membrane-displayed peptide ligand activates the pheromone response pathway in Saccharomyces cerevisiae, The Journal of Biochemistry, Volume 151, Issue 5, May 2012, Pages 551–557, https://doi.org/10.1093/jb/mvs027
Close - Share Icon Share
Abstract
The budding yeast, Saccharomyces cerevisiae, is an attractive host for studying G protein-coupled receptors (GPCRs). We developed a system in which a peptide ligand specific for GPCR is displayed on yeast plasma membrane. The model system described here is based on yeast plasma membrane display of an analogue of α-factor, which is a peptide ligand for Ste2p, the GPCR that activates the yeast pheromone response pathway. α-Factor analogues, containing linkers of varying lengths and produced in yeast cells, became attached to the cell plasma membrane by linking to the glycosylphosphatidylinositol (GPI)-anchored plasma membrane protein Yps1p. We were able to demonstrate that an optimized α-factor analogue activated the pheromone response pathway in S. cerevisiae, as assessed by a fluorescent reporter assay. Furthermore, it was shown that linker length strongly influenced signalling pathway activation. To our knowledge, this is the first report documenting functional signalling by a plasma membrane-displayed ligand in S. cerevisiae.
