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TitleEpoxidized graphene grid for highly efficient high-resolution cryoEM structural analysis.
Journal, issue, pagesSci Rep, Vol. 13, Issue 1, Page 2279, Year 2023
Publish dateFeb 8, 2023
AuthorsJunso Fujita / Fumiaki Makino / Haruyasu Asahara / Maiko Moriguchi / Shota Kumano / Itsuki Anzai / Jun-Ichi Kishikawa / Yoshiharu Matsuura / Takayuki Kato / Keiichi Namba / Tsuyoshi Inoue /
PubMed AbstractFunctionalization 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 ...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 as a mild oxidant and confirmed the oxidized graphene grid is storable with its functionality for at least three months under N 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 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.
External linksSci Rep / PubMed:36755111 / PubMed Central
MethodsEM (single particle)
Resolution1.29 - 3.23 Å
Structure data

EMDB-31310: GroEL on EG-grid
Method: EM (single particle) / Resolution: 1.99 Å

EMDB-31311: GroEL on hydrophilized graphene grid (low particle density)
Method: EM (single particle) / Resolution: 2.08 Å

EMDB-31312: A3B3 ring of V1-ATPase on EG-grid
Method: EM (single particle) / Resolution: 3.03 Å

EMDB-31313: Beta-galactosidase on EG-grid
Method: EM (single particle) / Resolution: 1.81 Å

EMDB-31314: Apoferritin on EG-grid
Method: EM (single particle) / Resolution: 1.29 Å

EMDB-32159: GroEL on hydrophilized graphene grid (high particle density)
Method: EM (single particle) / Resolution: 2.09 Å

EMDB-32160: SARS-CoV-2 spike protein (1-up RBD) on EG-grid
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-32161: SARS-CoV-2 spike protein (1-up RBD) on Quantifoil grid
Method: EM (single particle) / Resolution: 3.23 Å

EMDB-32162: GAPDH on EG-grid
Method: EM (single particle) / Resolution: 2.16 Å

EMDB-34106: GroEL on EG-grid stored for 3 months after graphene oxidation
Method: EM (single particle) / Resolution: 2.06 Å

EMDB-34743: GroEL on Quantifoil grid
Method: EM (single particle) / Resolution: 2.81 Å

Source
  • Escherichia coli (E. coli)
  • Escherichia coli
  • Thermus thermophilus (bacteria)
  • Homo sapiens (human)
  • Thermus thermophilus HB8 (bacteria)
  • Mus musculus (house mouse)
  • Mus musculu (house mouse)
  • Severe acute respiratory syndrome coronavirus 2

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