Cryo-EM analysis of Ebola virus nucleocapsid-like assembly

Yan Wang; Jennifer M Binning; Grigore D Pintilie; Wah Chiu; Gaya K Amarasinghe; Daisy W Leung; Zhaoming Su

doi:10.1016/j.xpro.2021.101030

Cryo-EM analysis of Ebola virus nucleocapsid-like assembly

STAR Protoc. 2021 Dec 16;3(1):101030. doi: 10.1016/j.xpro.2021.101030. eCollection 2022 Mar 18.

Authors

Yan Wang¹, Jennifer M Binning^{2

3}, Grigore D Pintilie^{4

5}, Wah Chiu^{4

5}, Gaya K Amarasinghe², Daisy W Leung^{2

6}, Zhaoming Su¹

Affiliations

¹ The State Key Laboratory of Biotherapy and Cancer Center, Department of Geriatrics and National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, Chengdu, Sichuan 610044, China.
² Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
³ Department of Molecular Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612, USA.
⁴ Department of Bioengineering and Department of Microbiology and Immunology, James H. Clark Center, Stanford University, Stanford, CA 94305, USA.
⁵ Division of Cryo-EM and Bioimaging, SSRL, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA.
⁶ Department of Medicine, Washington University School of Medicine, St. Louis, MO 63110, USA.

Abstract

This protocol describes the reconstitution of the filamentous Ebola virus nucleocapsid-like assembly in vitro. This is followed by solving the cryo-EM structure using helical reconstruction, and flexible fitting of the existing model into the 5.8 Å cryo-EM map. The protocol can be applied to other filamentous viral protein assemblies, particularly those with high flexibility and moderate resolution maps, which present technical challenges to model building. For complete details on the use and execution of this profile, please refer to Su et al. (2018).

Keywords: Cryo-EM; Microbiology; Protein Biochemistry; Protein expression and purification; Structural Biology.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Cryoelectron Microscopy / methods
Ebolavirus* / chemistry
Hemorrhagic Fever, Ebola*
Humans
Nucleocapsid / chemistry
Virus Assembly

Abstract

Publication types

MeSH terms

Grants and funding