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Uniform thin ice on ultraflat graphene for high-resolution cryo-EM

Cryo-electron microscopy (cryo-EM) visualizes the atomic structure of macromolecules that are embedded in vitrified thin ice at their close-to-native state. However, the homogeneity of ice thickness, a key factor to ensure high image quality, is poorly controlled during specimen preparation and has...

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Autores principales: Zheng, Liming, Liu, Nan, Gao, Xiaoyin, Zhu, Wenqing, Liu, Kun, Wu, Cang, Yan, Rui, Zhang, Jincan, Gao, Xin, Yao, Yating, Deng, Bing, Xu, Jie, Lu, Ye, Liu, Zhongmin, Li, Mengsen, Wei, Xiaoding, Wang, Hong-Wei, Peng, Hailin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group US 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9834055/
https://www.ncbi.nlm.nih.gov/pubmed/36522503
http://dx.doi.org/10.1038/s41592-022-01693-y
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author Zheng, Liming
Liu, Nan
Gao, Xiaoyin
Zhu, Wenqing
Liu, Kun
Wu, Cang
Yan, Rui
Zhang, Jincan
Gao, Xin
Yao, Yating
Deng, Bing
Xu, Jie
Lu, Ye
Liu, Zhongmin
Li, Mengsen
Wei, Xiaoding
Wang, Hong-Wei
Peng, Hailin
author_facet Zheng, Liming
Liu, Nan
Gao, Xiaoyin
Zhu, Wenqing
Liu, Kun
Wu, Cang
Yan, Rui
Zhang, Jincan
Gao, Xin
Yao, Yating
Deng, Bing
Xu, Jie
Lu, Ye
Liu, Zhongmin
Li, Mengsen
Wei, Xiaoding
Wang, Hong-Wei
Peng, Hailin
author_sort Zheng, Liming
collection PubMed
description Cryo-electron microscopy (cryo-EM) visualizes the atomic structure of macromolecules that are embedded in vitrified thin ice at their close-to-native state. However, the homogeneity of ice thickness, a key factor to ensure high image quality, is poorly controlled during specimen preparation and has become one of the main challenges for high-resolution cryo-EM. Here we found that the uniformity of thin ice relies on the surface flatness of the supporting film, and developed a method to use ultraflat graphene (UFG) as the support for cryo-EM specimen preparation to achieve better control of vitreous ice thickness. We show that the uniform thin ice on UFG improves the image quality of vitrified specimens. Using such a method we successfully determined the three-dimensional structures of hemoglobin (64 kDa), α-fetoprotein (67 kDa) with no symmetry, and streptavidin (52 kDa) at a resolution of 3.5 Å, 2.6 Å and 2.2 Å, respectively. Furthermore, our results demonstrate the potential of UFG for the fields of cryo-electron tomography and structure-based drug discovery.
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spelling pubmed-98340552023-01-13 Uniform thin ice on ultraflat graphene for high-resolution cryo-EM Zheng, Liming Liu, Nan Gao, Xiaoyin Zhu, Wenqing Liu, Kun Wu, Cang Yan, Rui Zhang, Jincan Gao, Xin Yao, Yating Deng, Bing Xu, Jie Lu, Ye Liu, Zhongmin Li, Mengsen Wei, Xiaoding Wang, Hong-Wei Peng, Hailin Nat Methods Article Cryo-electron microscopy (cryo-EM) visualizes the atomic structure of macromolecules that are embedded in vitrified thin ice at their close-to-native state. However, the homogeneity of ice thickness, a key factor to ensure high image quality, is poorly controlled during specimen preparation and has become one of the main challenges for high-resolution cryo-EM. Here we found that the uniformity of thin ice relies on the surface flatness of the supporting film, and developed a method to use ultraflat graphene (UFG) as the support for cryo-EM specimen preparation to achieve better control of vitreous ice thickness. We show that the uniform thin ice on UFG improves the image quality of vitrified specimens. Using such a method we successfully determined the three-dimensional structures of hemoglobin (64 kDa), α-fetoprotein (67 kDa) with no symmetry, and streptavidin (52 kDa) at a resolution of 3.5 Å, 2.6 Å and 2.2 Å, respectively. Furthermore, our results demonstrate the potential of UFG for the fields of cryo-electron tomography and structure-based drug discovery. Nature Publishing Group US 2022-12-15 2023 /pmc/articles/PMC9834055/ /pubmed/36522503 http://dx.doi.org/10.1038/s41592-022-01693-y Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Zheng, Liming
Liu, Nan
Gao, Xiaoyin
Zhu, Wenqing
Liu, Kun
Wu, Cang
Yan, Rui
Zhang, Jincan
Gao, Xin
Yao, Yating
Deng, Bing
Xu, Jie
Lu, Ye
Liu, Zhongmin
Li, Mengsen
Wei, Xiaoding
Wang, Hong-Wei
Peng, Hailin
Uniform thin ice on ultraflat graphene for high-resolution cryo-EM
title Uniform thin ice on ultraflat graphene for high-resolution cryo-EM
title_full Uniform thin ice on ultraflat graphene for high-resolution cryo-EM
title_fullStr Uniform thin ice on ultraflat graphene for high-resolution cryo-EM
title_full_unstemmed Uniform thin ice on ultraflat graphene for high-resolution cryo-EM
title_short Uniform thin ice on ultraflat graphene for high-resolution cryo-EM
title_sort uniform thin ice on ultraflat graphene for high-resolution cryo-em
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9834055/
https://www.ncbi.nlm.nih.gov/pubmed/36522503
http://dx.doi.org/10.1038/s41592-022-01693-y
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