A step-by-step protocol for capturing conformational snapshots of ligand gated ion channels by single-particle cryo-EM

Capturing conformational snapshots by single-particle cryo-EM facilitates the analysis of ligand binding and activation mechanisms for ion channels and other receptor complexes. Here, we present a protocol to capture intermediate states of nanodisc-reconstituted TRPV1. This protocol covers sample pr...

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Detalles Bibliográficos
Autores principales: Zhang, Kaihua, Julius, David, Cheng, Yifan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9530669/
https://www.ncbi.nlm.nih.gov/pubmed/36181684
http://dx.doi.org/10.1016/j.xpro.2022.101732
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author Zhang, Kaihua
Julius, David
Cheng, Yifan
author_facet Zhang, Kaihua
Julius, David
Cheng, Yifan
author_sort Zhang, Kaihua
collection PubMed
description Capturing conformational snapshots by single-particle cryo-EM facilitates the analysis of ligand binding and activation mechanisms for ion channels and other receptor complexes. Here, we present a protocol to capture intermediate states of nanodisc-reconstituted TRPV1. This protocol covers sample preparation, data acquisition, and image processing with focuses on the symmetry expansion and focused 3D classification. This protocol can be adapted to different proteins and samples. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2021).
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spelling pubmed-95306692022-10-05 A step-by-step protocol for capturing conformational snapshots of ligand gated ion channels by single-particle cryo-EM Zhang, Kaihua Julius, David Cheng, Yifan STAR Protoc Protocol Capturing conformational snapshots by single-particle cryo-EM facilitates the analysis of ligand binding and activation mechanisms for ion channels and other receptor complexes. Here, we present a protocol to capture intermediate states of nanodisc-reconstituted TRPV1. This protocol covers sample preparation, data acquisition, and image processing with focuses on the symmetry expansion and focused 3D classification. This protocol can be adapted to different proteins and samples. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2021). Elsevier 2022-09-30 /pmc/articles/PMC9530669/ /pubmed/36181684 http://dx.doi.org/10.1016/j.xpro.2022.101732 Text en © 2022 The Author(s) 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
Zhang, Kaihua
Julius, David
Cheng, Yifan
A step-by-step protocol for capturing conformational snapshots of ligand gated ion channels by single-particle cryo-EM
title A step-by-step protocol for capturing conformational snapshots of ligand gated ion channels by single-particle cryo-EM
title_full A step-by-step protocol for capturing conformational snapshots of ligand gated ion channels by single-particle cryo-EM
title_fullStr A step-by-step protocol for capturing conformational snapshots of ligand gated ion channels by single-particle cryo-EM
title_full_unstemmed A step-by-step protocol for capturing conformational snapshots of ligand gated ion channels by single-particle cryo-EM
title_short A step-by-step protocol for capturing conformational snapshots of ligand gated ion channels by single-particle cryo-EM
title_sort step-by-step protocol for capturing conformational snapshots of ligand gated ion channels by single-particle cryo-em
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9530669/
https://www.ncbi.nlm.nih.gov/pubmed/36181684
http://dx.doi.org/10.1016/j.xpro.2022.101732
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