Screening of stable G-protein-coupled receptor variants in Saccharomyces cerevisiae

Mitsunori Shiroishi, Takuya Kobayashi

研究成果: 書籍/レポート タイプへの寄稿

2 被引用数 (Scopus)


G-protein-coupled receptors (GPCRs) are not only the largest protein family, but as a whole, they represent the largest group of therapeutic drug targets. Recent successes in the determination of GPCR structures have relied on the stabilization of receptors to overcome the difficulties in expression and purification. Although a large quantity of purified protein is needed for structural determination, the majority of wild- type GPCRs are too unstable to express and purify on a large scale. Therefore, rapid screening of highly expressed stable receptor "variants" is crucial. It has been demonstrated that fusing green fluorescent protein (GFP) to a target membrane protein facilitates the evaluation of the physical properties of the membrane protein in detergent. Furthermore, the budding yeast Saccharomyces cerevisiae enables rapid construction of an expression vector via its own efficient homologous recombination system. Herein, we describe the protocols for rapid construction and screening of stable GPCR variants using GFP and S. cerevisiae.

ホスト出版物のタイトルStructural Proteomics
ホスト出版物のサブタイトルHigh-Throughput Methods: Second Edition
出版社Springer New York
出版ステータス出版済み - 12月 10 2014

!!!All Science Journal Classification (ASJC) codes

  • 生化学、遺伝学、分子生物学(全般)


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