|Title||Epoxidized graphene grid for highly efficient high-resolution cryoEM structural analysis.|
|Journal, issue, pages||Sci Rep, Vol. 13, Issue 1, Page 2279, Year 2023|
|Publish date||Feb 8, 2023|
|Authors||Junso Fujita / Fumiaki Makino / Haruyasu Asahara / Maiko Moriguchi / Shota Kumano / Itsuki Anzai / Jun-Ichi Kishikawa / Yoshiharu Matsuura / Takayuki Kato / Keiichi Namba / Tsuyoshi Inoue /|
|PubMed Abstract||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 ...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 links||Sci Rep / PubMed:36755111 / PubMed Central|
|Methods||EM (single particle)|
|Resolution||1.29 - 3.23 Å|
EMDB-31310: GroEL on EG-grid
EMDB-31311: GroEL on hydrophilized graphene grid (low particle density)
EMDB-31312: A3B3 ring of V1-ATPase on EG-grid
EMDB-31313: Beta-galactosidase on EG-grid
EMDB-31314: Apoferritin on EG-grid
EMDB-32159: GroEL on hydrophilized graphene grid (high particle density)
EMDB-32160: SARS-CoV-2 spike protein (1-up RBD) on EG-grid
EMDB-32161: SARS-CoV-2 spike protein (1-up RBD) on Quantifoil grid
EMDB-32162: GAPDH on EG-grid
EMDB-34106: GroEL on EG-grid stored for 3 months after graphene oxidation
EMDB-34743: GroEL on Quantifoil grid
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