Please wait a minute...
浙江大学学报（医学版）
浙江大学学报（医学版）  2019, Vol. 48 Issue (1): 39-43    DOI: 10.3785/j.issn.1008-9292.2019.02.07
专题报道     
单颗粒冷冻电镜揭开G蛋白偶联受体结构生物学研究的新篇章
李春桃1(),张会冰1,张岩1,2()
1. 浙江大学医学院生物物理系， 浙江 杭州 310058
2. 浙江大学医学院附属邵逸夫医院病理科， 浙江 杭州 310016
Single-particle cryo-electron microscopy opens new avenues in structural biology of G protein-coupled receptor
LI Chuntao1(),ZHANG Huibing1,ZHANG Yan1,2()
1.Department of Biophysics, Zhejiang University School of Medicine, Hangzhou 310058, China;
2.Department of Pathology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310016, China
 全文: PDF(1360 KB)   HTML( 22 )
摘要：

G蛋白偶联受体（GPCR）是生物体内一类庞大而又多样的细胞膜蛋白。GPCR可以应细胞外信号发生构象变化，进而结合不同效应蛋白激活下游多种信号转导通路，调控众多生命活动过程，参与几乎所有病理过程的发生和发展。近年来，冷冻电镜技术在研究生物大分子结构方面取得了突飞猛进的发展，基于冷冻电镜的GPCR信号转导复合物的高分辨率三维结构不断出现。本文阐述了GPCR和G蛋白复合物相互作用的共性结构特征——受体第六个跨膜螺旋的构象变化，也展示了GPCR识别不同G蛋白亚型选择性的结构基础。单颗粒冷冻电镜提供了更加高效地鉴定受体与配体相互作用分子机制的方法，为GPCR信号通路的机制研究以及基于结构的药物理性设计提供了重要信息。
关键词： 受体，G-蛋白偶联;  冷冻;  显微镜检查，电子;  GTP结合蛋白质类    
Abstract:

G protein-coupled receptors (GPCRs) represent the largest class of cell surface receptors, mediating wide range of cellular and physiological processes through their transducers, G proteins and the-arrestins participate in almost all pathological processes. Recent technological advances are revolutionizing the utility of cryo-electron microscopy (cryo-EM), leading to a tremendous progress in the structural studies of biological macromolecules and cryo-EM has played a leading role in the structural biology of GPCR signaling complex. New discoveries of high-resolution three-dimensional structures of GPCR signaling complexes based on cryo-EM have emerged vigorously, which depict the common structural characteristics of intermolecular interaction between GPCR and G protein complex-the conformational changes of the transmembrane helix 6 of receptors, and also demonstrate the structural basis of G protein subtype selectivity. Single-particle cryo-EM becomes an efficient tool for identifying the molecular mechanism of receptor-ligand interaction, providing important information for understanding GPCR signaling and the structure-based drug design.
Key words: Receptors, G-protein-coupled    Freezing    Microscopy, electron    GTP-binding proteins
收稿日期: 2018-09-30 出版日期: 2019-05-10
:  Q71  
基金资助: 浙江省自然科学基金(LR19H310001)
通讯作者: 张岩     E-mail: 21818621@zju.edu.cn;zhang_yan@zju.edu.cn
作者简介: 李春桃（1993—），女，硕士研究生，主要从事视紫红质类G蛋白偶联受体的结构与功能研究；E-mail： 21818621@zju.edu.cn；https://orcid.org/0000-0001-9802-5449|张　岩（1985—），男，博士，研究员，主要从事G蛋白偶联受体的结构与功能及其药理学研究；E-mail： zhang_yan@zju.edu.cn； https://orcid.org/0000-0003-2189-0244
服务　　
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章　　
李春桃
张会冰
张岩

引用本文:

李春桃,张会冰,张岩. 单颗粒冷冻电镜揭开G蛋白偶联受体结构生物学研究的新篇章[J]. 浙江大学学报（医学版）, 2019, 48(1): 39-43.

LI Chuntao,ZHANG Huibing,ZHANG Yan. Single-particle cryo-electron microscopy opens new avenues in structural biology of G protein-coupled receptor. J Zhejiang Univ (Med Sci), 2019, 48(1): 39-43.

链接本文:

http://www.zjujournals.com/med/CN/10.3785/j.issn.1008-9292.2019.02.07    或    http://www.zjujournals.com/med/CN/Y2019/V48/I1/39

图1  单颗粒冷冻电镜解析的G蛋白偶联受体复合物电镜密度图
1 HAUSER A S , CHAVALI S , MASUHO I , et al . Pharmacogenomics of GPCR drug targets[J]. Cell,2018,172(1-2):41-54.e19.
2 DEISENHOFER J , EPP O, MIKI K , et al . X-ray structure analysis of a membrane protein complex. Electron density map at 3 ? resolution and a model of the chromophores of the photosynthetic reaction center from Rhodopseudomonas viridis [J]. J Mol Biol,1984,180(2):385-398.
3 OKADA T , LE T I , FOX B A, et al . X-ray diffraction analysis of three-dimensional crystals of bovine rhodopsin obtained from mixed micelles[J]. J Struct Biol,2000,130(1):73-80.
4 GHOSH E , KUMARI P , JAIMAN D , et al . Methodological advances: the unsung heroes of the GPCR structural revolution[J]. Nat Rev Mol Cell Biol,2015,16(2):69-81.
5 RASMUSSEN S G , DEVREE B T , ZOU Y , et al . Crystal structure of the β2 adrenergic receptor-Gs protein complex[J]. Nature,2011，477(7366),549-555.
6 CARPENTER B , NEHMé R , WARNE T , et al . Structure of the adenosine A(2A) receptor bound to an engineered G protein[J]. Nature,2016,536(7614):104-107.
7 LIANG Y L , KHOSHOUEI M , RADJAINIA M , et al . Phase-plate cryo-EM structure of a class B GPCR-G-protein complex[J]. Nature,2017,546(7656):118-123.
8 ZHANG Y , SUN B , FENG D , et al . Cryo-EM structure of the activated GLP-1 receptor in complex with a G protein[J]. Nature,2017,546(7657):248-253.
9 LIANG Y L , KHOSHOUEI M , GLUKHOVA A , et al . Phase-plate cryo-EM structure of a biased agonist-bound human GLP-1 receptor-Gs complex[J]. Nature,2018,555(7694):121-125.
10 GARCíA-NAFRíA J , LEE Y, BAI X , et al . Cryo-EM structure of the adenosine A2A receptor coupled to an engineered heterotrimeric G protein[J/OL]. Elife,2018,7:e35946.
11 SAFDARI H A , PANDEY S , SHUKLA A K , et al . Illuminating GPCR Signaling by cryo-EM[J]. Trends Cell Biol,2018,28(8):591-594.
12 KANG Y , KUYBEDA O , WAAL P W DE , et al . Publisher correction: cryo-EM structure of human rhodopsin bound to an inhibitory G protein[J/OL]. Nature,2018,561(7724):E44.
13 KOEHL A , HU H , MAEDA S , et al . Structure of the μ-opioid receptor-Gi protein complex[J]. Nature,2018,558(7711):547-552.
14 DRAPER-JOYCE C J , KHOSHOUEI M , THAL D M , et al . Structure of the adenosine-bound human adenosine A1 receptor-Gi complex[J]. Nature,2018,558(7711):559-563.
15 GARCíA-NAFRíA J , NEHMé R , EDWARDS P C , et al . Cryo-EM structure of the serotonin 5-HT1B receptor coupled to heterotrimeric Go [J]. Nature,2018,558(7711):620-623.
16 KUMARI P , SRIVASTAVA A , BANERJEE R , et al . Functional competence of a partially engaged GPCR-beta-arrestin complex[J]. Nat Commun,2016,7:13416.
17 THOMSEN A R B , PLOUFFE B , CAHILL T J 3RD, et al . GPCR-G protein-β-arrestin super-complex mediates sustained G protein signaling[J]. Cell, 2016,166（4）:907-919.
18 CAHILL T J 3RD, THOMSEN A R , TARRASCH J T , et al . Distinct conformations of GPCR-beta-arrestin complexes mediate desensitization, signaling, and endocytosis[J]. Proc Natl Acad Sci U S A, 2017,114(10):2562-2567.
[1] 史婧,冯钰. 细菌RNA聚合酶抑制剂的分子生物学机制研究进展[J]. 浙江大学学报（医学版）, 2019, 48(1): 44-49.
[2] 张壮, 魏尔清, 卢韵碧. GPR17在中枢神经系统损伤修复中的作用研究进展[J]. 浙江大学学报（医学版）, 2013, 42(3): 355-359.
[3] 谭延斌，武新英，张景峰，张敏鸣. 超顺磁性氧化铁对血管内皮细胞的生物学影响及其磁共振成像效应[J]. 浙江大学学报（医学版）, 2010, 39(2): 118-124.
[4] 赵百亲;朱家光;章明;程海峰;范军强. 冷冻对心肌合成一氧化氮的影响[J]. 浙江大学学报（医学版）, 2005, 34(1): 77-79.
[5] 赵忠良, 吕新法, 吴健敏, 胡中荣. 冻融裂解和蛋白变性剂对肝组织内HBV抗原释放的影响[J]. 浙江大学学报（医学版）, 1993, 22(3): 115-116.
[6] 吕新法, 胡中荣, 王可宁, 徐晓, 朱智勇, 赵年丰, 马亦林. 流行性出血热病毒的冻融裂解及其机理探讨[J]. 浙江大学学报（医学版）, 1990, 19(2): 49-52.
[7] 王竞. 巩膜环扎伴冷凝术后的渗出性视网膜脱离[J]. 浙江大学学报（医学版）, 1988, 17(6): 267-268.