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

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

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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 ro-bust 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 mem-brane 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 sup-porting film in cryo-EM specimen preparation.

Reduced graphene oxideCryo-EM3D reconstructionAir-water interface

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

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Ministry of Education Key Laboratory of Protein Sciences,Beijing Advanced Innovation Center for Structural Bio-logy,School of Life Sciences,Tsinghua University,Beijing 100084,China

Tsinghua-Peking Joint Center for Life Sciences,Tsinghua University,Beijing 100084,China

Center for Nanochemistry,Beijing Science and Engineering Center for Nanocarbons,Beijing National Laboratory for Molecular Sciences,College of Chemistry and Molecular Engineering,Peking University,Beijing 100871,China

Joint Graduate Program of Peking-Tsinghua-NIBS,School of Life Sciences,Tsinghua University,Beijing 100084,China

Beijing Frontier Research Center for Biological Structures,Tsinghua University,Beijing 100084,China

Academy for Advanced Interdisciplinary Studies,Peking University,Beijing 100871,China

School of Materials Science and Engineering,Tianjin University,Tianjin 300072,China

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2016YFA050110031825009to H-W Wang215253102014CB932500 and 2016YFA0200101to H Peng

2021

生物物理学报
中国生物物理学会 中国科学院生物物理研究所

生物物理学报

影响因子：0.355
ISSN：1000-6737
年,卷(期)：2021.7(3)
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