Measuring Activation of Lipid G Protein- Coupled Receptors Using the TGF-á Shedding Assay

Asuka Inoue, Junken Aoki

研究成果: Chapter

抄録

Lipid mediators are intercellular bioactive lipid molecules that induce various cellular responses mostly through G protein-coupled receptors (GPCRs). According to the IUPHAR database and recent publications, there are now approximately 50 lipid-recognizing GPCRs. Monitoring activation of GPCRs by lipid mediators or synthetic ligands is crucial for studying the actions of bioactive lipids as well as lipid GPCRs. Because each GPCR differentially couples with heterotrimeric G proteins (G s, G i/o, G q/11, G 12/13), in general it is necessary to prepare multiple GPCR assays to detect distinct G-protein signaling and individual optimization. In this protocol section, we describe a recently developed transforming growth factor-á (TGF-á) shedding assay, which is a simple and accurate method for measuring activation of lipid GPCRs. By utilizing coexpression of chimeric Gá subunits, the TGF-á shedding assay can detect activation of a wide range of GPCRs that are coupled with any of the four G proteins. Indeed, 39 of 45 (87 %) lipid GPCRs examined were detectable in the single format of the TGF-á shedding assay, demonstrating, to the best of our knowledge, the greatest coverage of a lipid GPCR assay. Thus, the TGF-á shedding assay provides a useful platform for analyzing lipid GPCRs.

本文言語English
ホスト出版物のタイトルBioactive Lipid Mediators
ホスト出版物のサブタイトルCurrent Reviews and Protocols
出版社Springer Japan
ページ393-408
ページ数16
ISBN(電子版)9784431556695
ISBN(印刷版)9784431556688
DOI
出版ステータスPublished - 2015 1 1

ASJC Scopus subject areas

  • 生化学、遺伝学、分子生物学(全般)
  • 医学(全般)

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