DOI: 10.1073/pnas.2009385117
论文题名: Cryo-EM analysis of a membrane protein embedded in the liposome
作者: Yao X. ; Fan X. ; Yan N.
刊名: Proceedings of the National Academy of Sciences of the United States of America
ISSN: 0027-8424
出版年: 2020
卷: 117, 期: 31 起始页码: 18497
结束页码: 18503
语种: 英语
英文关键词: Cryo-em
; Graphene grids
; Membrane protein
; Proteoliposome
; Structural biology
Scopus关键词: bacterial protein
; graphene
; liposome
; membrane protein
; protein AcrB
; unclassified drug
; AcrB protein, E coli
; Escherichia coli protein
; liposome
; membrane protein
; multidrug resistance associated protein
; Article
; cryoelectron microscopy
; electrochemical analysis
; nonhuman
; priority journal
; protein analysis
; protein conformation
; protein function
; protein isolation
; three-dimensional imaging
; cell membrane
; cryoelectron microscopy
; Escherichia coli
; metabolism
; molecular model
; ultrastructure
; Cell Membrane
; Cryoelectron Microscopy
; Escherichia coli
; Escherichia coli Proteins
; Liposomes
; Membrane Proteins
; Models, Molecular
; Multidrug Resistance-Associated Proteins
; Protein Conformation
英文摘要: Membrane proteins (MPs) used to be the most difficult targets for structural biology when X-ray crystallography was the mainstream approach. With the resolution revolution of single-particle electron cryo-microscopy (cryo-EM), rapid progress has been made for structural elucidation of isolated MPs. The next challenge is to preserve the electrochemical gradients and membrane curvature for a comprehensive structural elucidation of MPs that rely on these chemical and physical properties for their biological functions. Toward this goal, here we present a convenient workflow for cryo-EM structural analysis of MPs embedded in liposomes, using the well-characterized AcrB as a prototype. Combining optimized proteoliposome isolation, cryo-sample preparation on graphene grids, and an efficient particle selection strategy, the threedimensional (3D) reconstruction of AcrB embedded in liposomes was obtained at 3.9 Å resolution. The conformation of the homotrimeric AcrB remains the same when the surrounding membranes display different curvatures. Our approach, which can be widely applied to cryo-EM analysis of MPs with distinctive soluble domains, lays out the foundation for cryo-EM analysis of integral or peripheral MPs whose functions are affected by transmembrane electrochemical gradients or/and membrane curvatures. © 2020 National Academy of Sciences. All rights reserved. Citation statistics:
资源类型: 期刊论文
标识符: http://119.78.100.158/handle/2HF3EXSE/163414
Appears in Collections: 气候变化与战略
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作者单位: Yao, X., Department of Molecular Biology, Princeton University, Princeton, NJ 08544, United States; Fan, X., Department of Molecular Biology, Princeton University, Princeton, NJ 08544, United States; Yan, N., Department of Molecular Biology, Princeton University, Princeton, NJ 08544, United States
Recommended Citation:
Yao X.,Fan X.,Yan N.. Cryo-EM analysis of a membrane protein embedded in the liposome[J]. Proceedings of the National Academy of Sciences of the United States of America,2020-01-01,117(31)