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Published on: June 13, 2021
Chimeric yeast G-protein α subunit harboring a 37-residue C-terminal gustducin-specific sequence is functional in
Keisuke Hara1, Yuko Inada, Takuya Ono
1Japan Society for the Promotion of Science, Sakyo-ku, Kyoto, Japan.
Abstract:
Despite many recent studies of G-protein-coupled receptor (GPCR) structures, it is not yet well understood how these receptors activate G proteins. The GPCR assay using baker's yeast, Saccharomyces cerevisiae, is an effective experimental model for the characterization of GPCR-Gα interactions. Here, using the yeast endogenous Gα protein (Gpa1p) as template, we constructed various chimeric Gα proteins with a region that is considered to be necessary for interaction with mammalian receptors. The signaling assay using the yeast pheromone receptor revealed that the chimeric Gα protein harboring 37 gustducin-specific amino acid residues at its C-terminus (GPA1/gust37) maintained functionality in yeast. In contrast, GPA1/gust44, a variant routinely used in mammalian experimental systems, was not functional.
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