Epoxidized graphene grid for highly efficient high-resolution cryoEM structural analysis
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 modification of graphene using photoactivated ClO2 · as a mild oxidant and confirmed the oxidi...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-02-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-29396-0 |
| _version_ | 1811165967214968832 |
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| author | Junso Fujita Fumiaki Makino Haruyasu Asahara Maiko Moriguchi Shota Kumano Itsuki Anzai Jun-ichi Kishikawa Yoshiharu Matsuura Takayuki Kato Keiichi Namba Tsuyoshi Inoue |
| author_facet | Junso Fujita Fumiaki Makino Haruyasu Asahara Maiko Moriguchi Shota Kumano Itsuki Anzai Jun-ichi Kishikawa Yoshiharu Matsuura Takayuki Kato Keiichi Namba Tsuyoshi Inoue |
| author_sort | Junso Fujita |
| collection | DOAJ |
| description | 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 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 N2 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. |
| first_indexed | 2024-04-10T15:44:45Z |
| format | Article |
| id | doaj.art-eaacd8fd1a16491b9ebd4322dcaea0e7 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-10T15:44:45Z |
| publishDate | 2023-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-eaacd8fd1a16491b9ebd4322dcaea0e72023-02-12T12:11:32ZengNature PortfolioScientific Reports2045-23222023-02-0113111510.1038/s41598-023-29396-0Epoxidized graphene grid for highly efficient high-resolution cryoEM structural analysisJunso Fujita0Fumiaki Makino1Haruyasu Asahara2Maiko Moriguchi3Shota Kumano4Itsuki Anzai5Jun-ichi Kishikawa6Yoshiharu Matsuura7Takayuki Kato8Keiichi Namba9Tsuyoshi Inoue10Graduate School of Frontier Biosciences, Osaka UniversityGraduate School of Frontier Biosciences, Osaka UniversityGraduate School of Pharmaceutical Sciences, Osaka UniversityGraduate School of Pharmaceutical Sciences, Osaka UniversityGraduate School of Pharmaceutical Sciences, Osaka UniversityDepartment of Molecular Virology, Research Institute for Microbial Diseases, Osaka UniversityInstitute for Protein Research, Osaka UniversityCenter for Infectious Disease Education and Research, Osaka UniversityInstitute for Protein Research, Osaka UniversityGraduate School of Frontier Biosciences, Osaka UniversityGraduate School of Pharmaceutical Sciences, Osaka UniversityAbstract 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 N2 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.https://doi.org/10.1038/s41598-023-29396-0 |
| spellingShingle | Junso Fujita Fumiaki Makino Haruyasu Asahara Maiko Moriguchi Shota Kumano Itsuki Anzai Jun-ichi Kishikawa Yoshiharu Matsuura Takayuki Kato Keiichi Namba Tsuyoshi Inoue Epoxidized graphene grid for highly efficient high-resolution cryoEM structural analysis Scientific Reports |
| title | Epoxidized graphene grid for highly efficient high-resolution cryoEM structural analysis |
| title_full | Epoxidized graphene grid for highly efficient high-resolution cryoEM structural analysis |
| title_fullStr | Epoxidized graphene grid for highly efficient high-resolution cryoEM structural analysis |
| title_full_unstemmed | Epoxidized graphene grid for highly efficient high-resolution cryoEM structural analysis |
| title_short | Epoxidized graphene grid for highly efficient high-resolution cryoEM structural analysis |
| title_sort | epoxidized graphene grid for highly efficient high resolution cryoem structural analysis |
| url | https://doi.org/10.1038/s41598-023-29396-0 |
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