Reduced graphene oxide membrane as supporting film for high-resolution cryo-EM

Nan Liu, Liming Zheng, Jie Xu, Jia Wang, Cuixia Hu, Jun Lan, Xing Zhang, Jincan Zhang, Kui Xu, Hang Cheng, Zi Yang, Xin Gao, Xinquan Wang, Hailin Peng, Yanan Chen, Hong Wei Wang

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

Although single-particle cryogenic electron microscopy (cryo-EM) has been applied extensively for elu-cidating many crucial biological mechanisms at the molecular level, this technique still faces critical challenges, the major one of which is to prepare the high-quality cryo-EM specimen. Aiming to achieve a more reproducible and efficient cryo-EM specimen preparation, novel supporting films including graphene-based two-dimensional materials have been explored in recent years. Here we report a robust and simple method to fabricate EM grids coated with single-or few-layer reduced graphene oxide (RGO) membrane in large batch for high-resolution cryo-EM structural determination. The RGO membrane has decreased interlayer space and enhanced electrical conductivity in comparison to regular graphene oxide (GO) membrane. Moreover, we found that the RGO supporting film exhibited nice particle-absorption ability, thus avoiding the air–water interface problem. More importantly, we found that the RGO supporting film is particularly useful in cryo-EM reconstruction of sub-100-kDa bio-molecules at near-atomic resolution, as exemplified by the study of RBD-ACE2 complex and other small protein molecules. We envision that the RGO membranes can be used as a robust graphene-based supporting film in cryo-EM specimen preparation.
Original languageEnglish (US)
Pages (from-to)227-238
Number of pages12
JournalBiophysics Reports
Volume7
Issue number3
DOIs
StatePublished - Jan 1 2021
Externally publishedYes

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Generated from Scopus record by KAUST IRTS on 2023-09-21

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