1. Chimeric Yeast G-Protein α Subunit Harboring a 37-Residue C-Terminal Gustducin-Specific Sequence Is Functional inSaccharomyces cerevisiae
- Author
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Masaji Ishiguro, Takaharu Tanaka, Mitsuyoshi Ueda, Yoshimi Yasuda-Kamatani, Keisuke Hara, Takuya Ono, Yuko Inada, Takumi Misaka, Keiko Abe, and Kouichi Kuroda
- Subjects
Models, Molecular ,Saccharomyces cerevisiae Proteins ,G protein ,Recombinant Fusion Proteins ,Molecular Sequence Data ,Saccharomyces cerevisiae ,Biology ,Applied Microbiology and Biotechnology ,Biochemistry ,Analytical Chemistry ,Amino Acid Sequence ,Transducin ,Receptor ,Molecular Biology ,Peptide sequence ,G protein-coupled receptor ,Organic Chemistry ,General Medicine ,biology.organism_classification ,Gustducin ,GTP-Binding Protein alpha Subunits ,Yeast ,Protein Structure, Tertiary ,G beta-gamma complex ,Genetic Engineering ,Biotechnology - 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.
- Published
- 2012
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