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- Title
Sub-3 Å Cryo-EM Structures of Necrosis Virus Particles via the Use of Multipurpose TEM with Electron Counting Camera.
- Authors
Wang, Chun-Hsiung; Chen, Dong-Hua; Huang, Shih-Hsin; Wu, Yi-Min; Chen, Yi-Yun; Hwu, Yeukuang; Bushnell, David; Kornberg, Roger; Chang, Wei-Hau
- Abstract
During this global pandemic, cryo-EM has made a great impact on the structure determination of COVID-19 proteins. However, nearly all high-resolution results are based on data acquired on state-of-the-art microscopes where their availability is restricted to a number of centers across the globe with the studies on infectious viruses being further regulated or forbidden. One potential remedy is to employ multipurpose microscopes. Here, we investigated the capability of 200 kV multipurpose microscopes equipped with a direct electron camera in determining the structures of infectious particles. We used 30 nm particles of the grouper nerve necrosis virus as a test sample and obtained the cryo-EM structure with a resolution as high as ∼2.7 Å from a setting that used electron counting. For comparison, we tested a high-end cryo-EM (Talos Arctica) using a similar virus (Macrobrachium rosenbergii nodavirus) to obtain virtually the same resolution. Those results revealed that the resolution is ultimately limited by the depth of field. Our work updates the density maps of these viruses at the sub-3Å level to allow for building accurate atomic models from de novo to provide structural insights into the assembly of the capsids. Importantly, this study demonstrated that multipurpose TEMs are capable of the high-resolution cryo-EM structure determination of infectious particles and is thus germane to the research on pandemics.
- Subjects
MACROBRACHIUM rosenbergii; ATOMIC models; ELECTRONS; CAMERAS; NECROSIS; PHOTON counting
- Publication
International Journal of Molecular Sciences, 2021, Vol 22, Issue 13, p6859
- ISSN
1661-6596
- Publication type
Article
- DOI
10.3390/ijms22136859