A genetically encoded small-size fluorescent pair reveals allosteric conformational changes of G proteins upon its interaction with GPCRs by fluorescence lifetime based FRET

Pan Shi; Yanan Zhang; Pei Lv; Wei Fang; Shenglong Ling; Xiaoqi Guo; Dong Li; Sanling Liu; Demeng Sun; Longhua Zhang; Dongsheng Liu; Ji-Shen Zheng; Changlin Tian

doi:10.1039/D0CC02691C

A genetically encoded small-size fluorescent pair reveals allosteric conformational changes of G proteins upon its interaction with GPCRs by fluorescence lifetime based FRET†

Pan Shi,‡*^a Yanan Zhang,‡^a Pei Lv,‡^a Wei Fang,^a Shenglong Ling,^a Xiaoqi Guo,^a Dong Li,^a Sanling Liu,^a Demeng Sun,^a Longhua Zhang,^a Dongsheng Liu,^b Ji-Shen Zheng

^a and Changlin Tian

*^acd

Author affiliations

* Corresponding authors

^a Hefei National Laboratory of Physical Science at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, P. R. China
E-mail: shipan@ustc.edu.cn, cltian@ustc.edu.cn

^b iHuman Institute, ShanghaiTech University, 99 HaiKe Road, Shanghai, China

^c High Magnetic Field Laboratory, Chinese Academy of Sciences, Hefei, China

^d Anhui Laboratory of Advanced Photon Science and Technology, University of Science and Technology of China, Hefei 230026, P. R. China

Abstract

The dynamics of GPCRs (G protein-coupled receptors) coupling for cognate G proteins play a critical role in signal transduction. Herein, we reported a site-specifically labelled small-sized fluorescent pair 7-HC/FlAsH ((7-hydroxycoumarin-4-yl)-ethylglycine/fluorescein arsenical hairpin) for fluorescence lifetime based FRET (fluorescence resonance energy transfer) to reveal conformational differences of Gα_i1 (inhibitory G proteins) and Gα_s (stimulatory G proteins) upon β₂AR (β₂-adrenergic receptor) coupling. It offers a new generally applicable method to probe protein dynamic interactions or conformational changes.

Chemical Communications

A genetically encoded small-size fluorescent pair reveals allosteric conformational changes of G proteins upon its interaction with GPCRs by fluorescence lifetime based FRET†

Abstract

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A genetically encoded small-size fluorescent pair reveals allosteric conformational changes of G proteins upon its interaction with GPCRs by fluorescence lifetime based FRET

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