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

Mitsunori Shiroishi, Takuya Kobayashi

研究成果: 著書/レポートタイプへの貢献

2 引用 (Scopus)

抄録

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.

元の言語英語
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
出版者Springer New York
ページ159-170
ページ数12
ISBN(電子版)9781493922307
ISBN(印刷物)9781493922291
DOI
出版物ステータス出版済み - 12 10 2014

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G-Protein-Coupled Receptors
Yeast
Saccharomyces cerevisiae
Screening
Green Fluorescent Proteins
Membrane Proteins
Saccharomycetales
Homologous Recombination
Protein S
Detergents
Purification
Proteins
Stabilization
Physical properties
Pharmaceutical Preparations
Therapeutics

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

これを引用

Shiroishi, M., & Kobayashi, T. (2014). Screening of stable G-protein-coupled receptor variants in Saccharomyces cerevisiae. : Structural Proteomics: High-Throughput Methods: Second Edition (pp. 159-170). Springer New York. https://doi.org/10.1007/978-1-4939-2230-7_9

Screening of stable G-protein-coupled receptor variants in Saccharomyces cerevisiae. / Shiroishi, Mitsunori; Kobayashi, Takuya.

Structural Proteomics: High-Throughput Methods: Second Edition. Springer New York, 2014. p. 159-170.

研究成果: 著書/レポートタイプへの貢献

Shiroishi, M & Kobayashi, T 2014, Screening of stable G-protein-coupled receptor variants in Saccharomyces cerevisiae. : Structural Proteomics: High-Throughput Methods: Second Edition. Springer New York, pp. 159-170. https://doi.org/10.1007/978-1-4939-2230-7_9
Shiroishi M, Kobayashi T. Screening of stable G-protein-coupled receptor variants in Saccharomyces cerevisiae. : Structural Proteomics: High-Throughput Methods: Second Edition. Springer New York. 2014. p. 159-170 https://doi.org/10.1007/978-1-4939-2230-7_9
Shiroishi, Mitsunori ; Kobayashi, Takuya. / Screening of stable G-protein-coupled receptor variants in Saccharomyces cerevisiae. Structural Proteomics: High-Throughput Methods: Second Edition. Springer New York, 2014. pp. 159-170
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