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

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,...

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Detalles Bibliográficos
Autores principales: Wang, Wenshuai, Fedorova, Olga, Pyle, Anna Marie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10026036/
https://www.ncbi.nlm.nih.gov/pubmed/36920909
http://dx.doi.org/10.1016/j.xpro.2023.102166
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author Wang, Wenshuai
Fedorova, Olga
Pyle, Anna Marie
author_facet Wang, Wenshuai
Fedorova, Olga
Pyle, Anna Marie
author_sort Wang, Wenshuai
collection PubMed
description 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)
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spelling pubmed-100260362023-03-21 A protocol for capturing RNA-sensing innate immune receptors in multiple conformations by single-particle cryo-EM Wang, Wenshuai Fedorova, Olga Pyle, Anna Marie STAR Protoc Protocol 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) Elsevier 2023-03-14 /pmc/articles/PMC10026036/ /pubmed/36920909 http://dx.doi.org/10.1016/j.xpro.2023.102166 Text en © 2023 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Wang, Wenshuai
Fedorova, Olga
Pyle, Anna Marie
A protocol for capturing RNA-sensing innate immune receptors in multiple conformations by single-particle cryo-EM
title A protocol for capturing RNA-sensing innate immune receptors in multiple conformations by single-particle cryo-EM
title_full A protocol for capturing RNA-sensing innate immune receptors in multiple conformations by single-particle cryo-EM
title_fullStr A protocol for capturing RNA-sensing innate immune receptors in multiple conformations by single-particle cryo-EM
title_full_unstemmed A protocol for capturing RNA-sensing innate immune receptors in multiple conformations by single-particle cryo-EM
title_short A protocol for capturing RNA-sensing innate immune receptors in multiple conformations by single-particle cryo-EM
title_sort protocol for capturing rna-sensing innate immune receptors in multiple conformations by single-particle cryo-em
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10026036/
https://www.ncbi.nlm.nih.gov/pubmed/36920909
http://dx.doi.org/10.1016/j.xpro.2023.102166
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