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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,...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2023
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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) |
format | Online Article Text |
id | pubmed-10026036 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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