Chimeric Yeast G-Protein α Subunit Harboring a 37-Residue C-Terminal Gustducin-Specific Sequence Is Functional in Saccharomyces cerevisiae

Keisuke HARA; Yuko INADA; Takuya ONO; Kouichi KURODA; Yoshimi YASUDA-KAMATANI; Masaji ISHIGURO; Takaharu TANAKA; Takumi MISAKA; Keiko ABE; Mitsuyoshi UEDA

doi:10.1271/bbb.110820

Biochemistry & Molecular Biology Regular Papers

Chimeric Yeast G-Protein α Subunit Harboring a 37-Residue C-Terminal Gustducin-Specific Sequence Is Functional in Saccharomyces cerevisiae

Keisuke HARA, Yuko INADA, Takuya ONO, Kouichi KURODA, Yoshimi YASUDA-KAMATANI, Masaji ISHIGURO, Takaharu TANAKA, Takumi MISAKA, Keiko ABE, Mitsuyoshi UEDA

Author information

Keisuke HARA
Japan Society for the Promotion of Science
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University
Yuko INADA
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University
Takuya ONO
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University
Kouichi KURODA
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University
Yoshimi YASUDA-KAMATANI
Suntory Institute for Bioorganic Research
Masaji ISHIGURO
Suntory Institute for Bioorganic Research
Niigata University of Pharmacy and Applied Life Sciences
Takaharu TANAKA
Suntory Institute for Bioorganic Research
Takumi MISAKA
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo
Keiko ABE
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo
Mitsuyoshi UEDA
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University

Keywords: yeast G-protein-coupled receptor (GPCR) assay, chimeric Gα, G-protein-coupled receptor (GPCR)-Gα interaction, gustducin

JOURNAL FREE ACCESS

2012 Volume 76 Issue 3 Pages 512-516

DOI https://doi.org/10.1271/bbb.110820

View "Advance Publication" version (March 7, 2012).

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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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