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-Structure paper
タイトル | Uniform thin ice on ultraflat graphene for high-resolution cryo-EM. |
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ジャーナル・号・ページ | Nat Methods, Vol. 20, Issue 1, Page 123-130, Year 2023 |
掲載日 | 2022年12月15日 |
著者 | Liming Zheng / Nan Liu / Xiaoyin Gao / Wenqing Zhu / Kun Liu / Cang Wu / Rui Yan / Jincan Zhang / Xin Gao / Yating Yao / Bing Deng / Jie Xu / Ye Lu / Zhongmin Liu / Mengsen Li / Xiaoding Wei / Hong-Wei Wang / Hailin Peng / |
PubMed 要旨 | 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, ...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. |
リンク | Nat Methods / PubMed:36522503 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 2.2 - 3.5 Å |
構造データ | EMDB-33189, PDB-7xgy: EMDB-33861, PDB-7yim: PDB-8gvk: |
化合物 | ChemComp-HEM: ChemComp-HOH: |
由来 |
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キーワード | OXYGEN TRANSPORT / tetramer / METAL BINDING PROTEIN / metal binding / fatty acids binding / PROTEIN BINDING / biotin-binding protein |