In vitro expression and analysis of the 826 human G protein-coupled receptors.

Xuechen LV; Junlin Liu; Qiaoyun Shi; Qiwen Tan; Dong WU; John J Skinner; Angela L Walker; Lixia Zhao; Xiangxiang GU; Na Chen; Lu Xue; Pei SI; Lu Zhang; Zeshi Wang; Vsevolod Katritch; Zhi-jie Liu; Raymond C Stevens

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In vitro expression and analysis of the 826 human G protein-coupled receptors.

Author: Xuechen LV ¹ ; Junlin LIU ¹ ; Qiaoyun SHI ¹ ; Qiwen TAN ¹ ; Dong WU ¹ ; John J SKINNER ¹ ; Angela L WALKER ² ; Lixia ZHAO ¹ ; Xiangxiang GU ¹ ; Na CHEN ¹ ; Lu XUE ¹ ; Pei SI ¹ ; Lu ZHANG ¹ ; Zeshi WANG ¹ ; Vsevolod KATRITCH ² ; Zhi-Jie LIU ¹ ; Raymond C STEVENS ³
Author Information

1. iHuman Institute, ShanghaiTech University, Shanghai, 201210, China.
2. Department of Biological Sciences, Bridge Institute, University of Southern California, Los Angeles, CA, 90089, USA.
3. iHuman Institute, ShanghaiTech University, Shanghai, 201210, China. stevens@shanghaitech.edu.cn.
Publication Type:Journal Article
Keywords: G protein-coupled receptors; fusion construct; insect; protein expression; surface expression analysis
MeSH: Animals; Computational Biology; Crystallography, X-Ray; Gene Expression; Humans; Plasmids; genetics; metabolism; Protein Domains; Receptors, Adrenergic, beta-1; Receptors, G-Protein-Coupled; classification; genetics; metabolism; Receptors, Odorant; metabolism; Receptors, Purinergic P1; genetics; metabolism; Sf9 Cells; Spodoptera
From: Protein & Cell 2016;7(5):325-337
CountryChina
Language:English
Abstract: G protein-coupled receptors (GPCRs) are involved in all human physiological systems where they are responsible for transducing extracellular signals into cells. GPCRs signal in response to a diverse array of stimuli including light, hormones, and lipids, where these signals affect downstream cascades to impact both health and disease states. Yet, despite their importance as therapeutic targets, detailed molecular structures of only 30 GPCRs have been determined to date. A key challenge to their structure determination is adequate protein expression. Here we report the quantification of protein expression in an insect cell expression system for all 826 human GPCRs using two different fusion constructs. Expression characteristics are analyzed in aggregate and among each of the five distinct subfamilies. These data can be used to identify trends related to GPCR expression between different fusion constructs and between different GPCR families, and to prioritize lead candidates for future structure determination feasibility.