• 文献标题:   Epoxidized graphene grid for highly efficient high-resolution cryoEM structural analysis
  • 文献类型:   Article
  • 作  者:   FUJITA J, MAKINO F, ASAHARA H, MORIGUCHI M, KUMANO S, ANZAI I, KISHIKAWA J, MATSUURA Y, KATO T, NAMBA K, INOUE T
  • 作者关键词:  
  • 出版物名称:   SCIENTIFIC REPORTS
  • ISSN:   2045-2322
  • 通讯作者地址:  
  • 被引频次:   0
  • DOI:   10.1038/s41598-023-29396-0
  • 出版年:   2023

▎ 摘  要

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 ClO2 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 (TM)), 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 angstrom resolution from only 504 and 241 micrographs, respectively. A sample solution of 0.1 mg ml(-1) was sufficient to reconstruct a 3.10 angstrom resolution map of SARS-CoV-2 spike protein from 1163 micrographs. The map resolutions of beta-galactosidase and apoferritin easily reached 1.81 angstrom and 1.29 angstrom 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.