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使用ONE-GO生物传感器研究G蛋白偶联受体信号动力学和浓度依赖性反应的方案。

Protocol to investigate G protein-coupled receptor signaling kinetics and concentration-dependent responses using ONE-GO biosensors.

作者信息

Janicot Remi, Garcia-Marcos Mikel

机构信息

Department of Biochemistry & Cell Biology, Chobanian & Avedisian School of Medicine, Boston University, Boston, MA 02118, USA.

Department of Biochemistry & Cell Biology, Chobanian & Avedisian School of Medicine, Boston University, Boston, MA 02118, USA; Department of Biology, College of Arts & Sciences, Boston University, Boston, MA 02115, USA.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103383. doi: 10.1016/j.xpro.2024.103383. Epub 2024 Oct 11.

Abstract

ONE vector G protein optical (ONE-GO) biosensors are versatile tools to measure the activity of G protein-coupled receptors (GPCRs) in cells. The availability of ONE-GO biosensors for ten active Gα subunits representative of all four G protein families (G, G, G, and G) permits the study of virtually any GPCR. Here, we present a protocol to implement ONE-GO biosensors in cell lines to investigate GPCR signaling kinetics and concentration-dependent responses. We describe steps for cell culture and transfection, response measurement, and data analysis. For complete details on the use and execution of this protocol, please refer to Janicot et al..

摘要

单向量G蛋白光学(ONE-GO)生物传感器是用于测量细胞中G蛋白偶联受体(GPCR)活性的多功能工具。针对代表所有四个G蛋白家族(Gαs、Gαi/o、Gαq/11和Gα12/13)的十个活性Gα亚基的ONE-GO生物传感器,使得几乎任何GPCR的研究都成为可能。在这里,我们展示了一种在细胞系中应用ONE-GO生物传感器来研究GPCR信号动力学和浓度依赖性反应的方案。我们描述了细胞培养和转染、反应测量及数据分析的步骤。有关本方案使用和执行的完整详细信息,请参考贾尼科特等人的研究。

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