Work

A protocol for capturing RNA-sensing innate immune receptors in multiple conformations by single-particle cryo-EM

Year: 2023

Type: article

Abstract: Capturing different conformations of receptor proteins that are complexed with ligands by single-particle cryo-EM facilitates our understanding toward the mechanisms of ligand recognition and receptor activation cascades. Here, we present a protocol for capturing RNA-sensing innate immune receptors, such as RIG-I, in multiple conformations by single-particle cryo-EM. We describe steps for protein-ligand sample preparation, data acquisition, and image processing covering focused three-dimensional classification. This protocol can be adapted to capture the dynamic behavior of other receptors that can be stabilized. For complete details on the use and execution of this protocol, please refer to Wang and Pyle (2022).1

Source: STAR Protocols

DOI: 10.1016/j.xpro.2023.102166

Authors: Wenshuai Wang, Olga Fedorova, Anna Marie Pyle

Institutions: Howard Hughes Medical Institute, Yale University

Language: English

Topic: Advanced Electron Microscopy Techniques and Applications

Subfield: Structural Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $1,500

Funders: Howard Hughes Medical Institute, Yale University, National Institutes of Health, National Institute of Allergy and Infectious Diseases