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

STAR Protocols (Mar 2022)

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

Affiliations

Yan Wang: 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
Jennifer M. Binning: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA; Department of Molecular Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612, USA
Grigore D. Pintilie: Department of Bioengineering and Department of Microbiology and Immunology, James H. Clark Center, Stanford University, Stanford, CA 94305, USA; Division of Cryo-EM and Bioimaging, SSRL, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA
Wah Chiu: Department of Bioengineering and Department of Microbiology and Immunology, James H. Clark Center, Stanford University, Stanford, CA 94305, USA; Division of Cryo-EM and Bioimaging, SSRL, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA
Gaya K. Amarasinghe: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA; Corresponding author
Daisy W. Leung: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA; Department of Medicine, Washington University School of Medicine, St. Louis, MO 63110, USA; Corresponding author
Zhaoming Su: 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; Corresponding author

Journal volume & issue: Vol. 3, no. 1
p. 101030

Abstract

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Summary: 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).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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