Purification and cryo-EM structure determination of Arabidopsis thaliana GLR3.4

Ionotropic glutamate receptors (iGluRs) are ligand-gated ion channels that play crucial roles in the central nervous system. iGluR homologs, termed glutamate receptor-like channels (GLRs), have been found in plants. Investigating the structural and functional relationship between iGluRs and GLRs was...

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Autores principales: Gangwar, Shanti Pal, Green, Marriah N., Yelshanskaya, Maria V., Sobolevsky, Alexander I.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8496305/
https://www.ncbi.nlm.nih.gov/pubmed/34647037
http://dx.doi.org/10.1016/j.xpro.2021.100855
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author Gangwar, Shanti Pal
Green, Marriah N.
Yelshanskaya, Maria V.
Sobolevsky, Alexander I.
author_facet Gangwar, Shanti Pal
Green, Marriah N.
Yelshanskaya, Maria V.
Sobolevsky, Alexander I.
author_sort Gangwar, Shanti Pal
collection PubMed
description Ionotropic glutamate receptors (iGluRs) are ligand-gated ion channels that play crucial roles in the central nervous system. iGluR homologs, termed glutamate receptor-like channels (GLRs), have been found in plants. Investigating the structural and functional relationship between iGluRs and GLRs was limited by GLR protein expression, purification, and structural characterization. Here, we provide a detailed protocol for Arabidopsis thaliana GLR3.4 (AtGLR3.4) expression in a mammalian cell line and purification for structure determination by cryogenic electron microscopy (cryo-EM). For the complete details on the use and execution of this protocol, please refer to Green et al. (2021).
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spelling pubmed-84963052021-10-12 Purification and cryo-EM structure determination of Arabidopsis thaliana GLR3.4 Gangwar, Shanti Pal Green, Marriah N. Yelshanskaya, Maria V. Sobolevsky, Alexander I. STAR Protoc Protocol Ionotropic glutamate receptors (iGluRs) are ligand-gated ion channels that play crucial roles in the central nervous system. iGluR homologs, termed glutamate receptor-like channels (GLRs), have been found in plants. Investigating the structural and functional relationship between iGluRs and GLRs was limited by GLR protein expression, purification, and structural characterization. Here, we provide a detailed protocol for Arabidopsis thaliana GLR3.4 (AtGLR3.4) expression in a mammalian cell line and purification for structure determination by cryogenic electron microscopy (cryo-EM). For the complete details on the use and execution of this protocol, please refer to Green et al. (2021). Elsevier 2021-10-01 /pmc/articles/PMC8496305/ /pubmed/34647037 http://dx.doi.org/10.1016/j.xpro.2021.100855 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Gangwar, Shanti Pal
Green, Marriah N.
Yelshanskaya, Maria V.
Sobolevsky, Alexander I.
Purification and cryo-EM structure determination of Arabidopsis thaliana GLR3.4
title Purification and cryo-EM structure determination of Arabidopsis thaliana GLR3.4
title_full Purification and cryo-EM structure determination of Arabidopsis thaliana GLR3.4
title_fullStr Purification and cryo-EM structure determination of Arabidopsis thaliana GLR3.4
title_full_unstemmed Purification and cryo-EM structure determination of Arabidopsis thaliana GLR3.4
title_short Purification and cryo-EM structure determination of Arabidopsis thaliana GLR3.4
title_sort purification and cryo-em structure determination of arabidopsis thaliana glr3.4
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8496305/
https://www.ncbi.nlm.nih.gov/pubmed/34647037
http://dx.doi.org/10.1016/j.xpro.2021.100855
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