| Title: | Chimeric yeast G-protein alpha subunit harboring a 37-residue C-terminal gustducin-specific sequence is functional in Saccharomyces cerevisiae |
| Author(s): | Hara K; Inada Y; Ono T; Kuroda K; Yasuda-Kamatani Y; Ishiguro M; Tanaka T; Misaka T; Abe K; Ueda M; |
| Address: | "Japan Society for the Promotion of Science, Sakyo-ku, Kyoto, Japan" |
| Journal Title: | Biosci Biotechnol Biochem |
| ISSN/ISBN: | 1347-6947 (Electronic) 0916-8451 (Linking) |
| 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-Galpha interactions. Here, using the yeast endogenous Galpha protein (Gpa1p) as template, we constructed various chimeric Galpha 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 Galpha 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" |
| Keywords: | "Amino Acid Sequence GTP-Binding Protein alpha Subunits/chemistry/genetics/*metabolism Genetic Engineering/*methods Models, Molecular Molecular Sequence Data Protein Structure, Tertiary Recombinant Fusion Proteins/chemistry/genetics/*metabolism Saccharomyc;" |
| Notes: | "MedlineHara, Keisuke Inada, Yuko Ono, Takuya Kuroda, Kouichi Yasuda-Kamatani, Yoshimi Ishiguro, Masaji Tanaka, Takaharu Misaka, Takumi Abe, Keiko Ueda, Mitsuyoshi eng Research Support, Non-U.S. Gov't England 2012/03/28 Biosci Biotechnol Biochem. 2012; 76(3):512-6. doi: 10.1271/bbb.110820" |
