Epoxidized graphene grid for highly efficient high-resolution cryoEM structural analysis

Functionalization of graphene is one of the most important fundamental technologies in a wide variety of fields including industry and biochemistry. We have successfully achieved a novel oxidative modification of graphene using photoactivated ClO 2 · as a mild oxidant and confirmed the oxidized grap...

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Bibliographic Details
Published in:Scientific reports 2023-02, Vol.13 (1), p.2279-2279, Article 2279
Main Authors: Fujita, Junso, Makino, Fumiaki, Asahara, Haruyasu, Moriguchi, Maiko, Kumano, Shota, Anzai, Itsuki, Kishikawa, Jun-ichi, Matsuura, Yoshiharu, Kato, Takayuki, Namba, Keiichi, Inoue, Tsuyoshi
Format: Article
Language:English
Subjects:
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631/535/1258
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639/638
639/638/439
Apoferritin
Biological analysis
COVID-19
Cryoelectron Microscopy - methods
Glyceraldehyde
Glyceraldehyde 3-phosphate
Glyceraldehyde-3-phosphate dehydrogenase
Graphene
Graphite
Humanities and Social Sciences
Humans
Image processing
Macromolecules
Micrography
multidisciplinary
Oxidants
Oxidizing agents
Proteins
SARS-CoV-2
Science
Science (multidisciplinary)
Severe acute respiratory syndrome coronavirus 2
Spike protein
Structural analysis
β-Galactosidase
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Summary:Functionalization of graphene is one of the most important fundamental technologies in a wide variety of fields including industry and biochemistry. We have successfully achieved a novel oxidative modification of graphene using photoactivated ClO 2 · as a mild oxidant and confirmed the oxidized graphene grid is storable with its functionality for at least three months under N 2 atmosphere. Subsequent chemical functionalization enabled us to develop an epoxidized graphene grid (EG-grid™), which effectively adsorbs protein particles for electron cryomicroscopy (cryoEM) image analysis. The EG-grid dramatically improved the particle density and orientation distribution. The density maps of GroEL and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were reconstructed at 1.99 and 2.16 Å resolution from only 504 and 241 micrographs, respectively. A sample solution of 0.1 mg ml −1 was sufficient to reconstruct a 3.10 Å resolution map of SARS-CoV-2 spike protein from 1163 micrographs. The map resolutions of β-galactosidase and apoferritin easily reached 1.81 Å and 1.29 Å resolution, respectively, indicating its atomic-resolution imaging capability. Thus, the EG-grid will be an extremely powerful tool for highly efficient high-resolution cryoEM structural analysis of biological macromolecules.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-023-29396-0