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Self-assembled superstructure alleviates air-water interface effect in cryo-EM

Authors :
Liming Zheng
Jie Xu
Weihua Wang
Xiaoyin Gao
Chao Zhao
Weijun Guo
Luzhao Sun
Hang Cheng
Fanhao Meng
Buhang Chen
Weiyu Sun
Xia Jia
Xiong Zhou
Kai Wu
Zhongfan Liu
Feng Ding
Nan Liu
Hong-Wei Wang
Hailin Peng
Source :
Nature Communications, Vol 15, Iss 1, Pp 1-10 (2024)
Publication Year :
2024
Publisher :
Nature Portfolio, 2024.

Abstract

Abstract Cryo-electron microscopy (cryo-EM) has been widely used to reveal the structures of proteins at atomic resolution. One key challenge is that almost all proteins are predominantly adsorbed to the air-water interface during standard cryo-EM specimen preparation. The interaction of proteins with air-water interface will significantly impede the success of reconstruction and achievable resolution. Here, we highlight the critical role of impenetrable surfactant monolayers in passivating the air-water interface problems, and develop a robust effective method for high-resolution cryo-EM analysis, by using the superstructure GSAMs which comprises surfactant self-assembled monolayers (SAMs) and graphene membrane. The GSAMs works well in enriching the orientations and improving particle utilization ratio of multiple proteins, facilitating the 3.3-Å resolution reconstruction of a 100-kDa protein complex (ACE2-RBD), which shows strong preferential orientation using traditional specimen preparation protocol. Additionally, we demonstrate that GSAMs enables the successful determinations of small proteins (

Subjects

Subjects :
Science

Details

Language :
English
ISSN :
20411723
Volume :
15
Issue :
1
Database :
Directory of Open Access Journals
Journal :
Nature Communications
Publication Type :
Academic Journal
Accession number :
edsdoj.09347e7f08444754a630ab622f8e4178
Document Type :
article
Full Text :
https://doi.org/10.1038/s41467-024-51696-w