HDX-guided EPR spectroscopy to interrogate membrane protein dynamics

Solvent accessibilities of and distances between protein residues measured by pulsed-EPR approaches provide high-resolution information on dynamic protein motions. We describe protocols for the purification and site-directed spin labeling of integral membrane proteins. In our protocol, peptide-level...

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Detalles Bibliográficos
Autores principales: Lane, Benjamin J., Wang, Bolin, Ma, Yue, Calabrese, Antonio N., El Mkami, Hassane, Pliotas, Christos
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9304679/
https://www.ncbi.nlm.nih.gov/pubmed/35874470
http://dx.doi.org/10.1016/j.xpro.2022.101562
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author Lane, Benjamin J.
Wang, Bolin
Ma, Yue
Calabrese, Antonio N.
El Mkami, Hassane
Pliotas, Christos
author_facet Lane, Benjamin J.
Wang, Bolin
Ma, Yue
Calabrese, Antonio N.
El Mkami, Hassane
Pliotas, Christos
author_sort Lane, Benjamin J.
collection PubMed
description Solvent accessibilities of and distances between protein residues measured by pulsed-EPR approaches provide high-resolution information on dynamic protein motions. We describe protocols for the purification and site-directed spin labeling of integral membrane proteins. In our protocol, peptide-level HDX-MS is used as a precursor to guide single-residue resolution ESEEM accessibility measurements and spin labeling strategies for EPR applications. Exploiting the pentameric MscL channel as a model, we discuss the use of cwEPR, DEER/PELDOR, and ESEEM spectroscopies to interrogate membrane protein dynamics. For complete details on the use and execution of this protocol, please refer to Wang et al. (2022).
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spelling pubmed-93046792022-07-23 HDX-guided EPR spectroscopy to interrogate membrane protein dynamics Lane, Benjamin J. Wang, Bolin Ma, Yue Calabrese, Antonio N. El Mkami, Hassane Pliotas, Christos STAR Protoc Protocol Solvent accessibilities of and distances between protein residues measured by pulsed-EPR approaches provide high-resolution information on dynamic protein motions. We describe protocols for the purification and site-directed spin labeling of integral membrane proteins. In our protocol, peptide-level HDX-MS is used as a precursor to guide single-residue resolution ESEEM accessibility measurements and spin labeling strategies for EPR applications. Exploiting the pentameric MscL channel as a model, we discuss the use of cwEPR, DEER/PELDOR, and ESEEM spectroscopies to interrogate membrane protein dynamics. For complete details on the use and execution of this protocol, please refer to Wang et al. (2022). Elsevier 2022-07-18 /pmc/articles/PMC9304679/ /pubmed/35874470 http://dx.doi.org/10.1016/j.xpro.2022.101562 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
Lane, Benjamin J.
Wang, Bolin
Ma, Yue
Calabrese, Antonio N.
El Mkami, Hassane
Pliotas, Christos
HDX-guided EPR spectroscopy to interrogate membrane protein dynamics
title HDX-guided EPR spectroscopy to interrogate membrane protein dynamics
title_full HDX-guided EPR spectroscopy to interrogate membrane protein dynamics
title_fullStr HDX-guided EPR spectroscopy to interrogate membrane protein dynamics
title_full_unstemmed HDX-guided EPR spectroscopy to interrogate membrane protein dynamics
title_short HDX-guided EPR spectroscopy to interrogate membrane protein dynamics
title_sort hdx-guided epr spectroscopy to interrogate membrane protein dynamics
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9304679/
https://www.ncbi.nlm.nih.gov/pubmed/35874470
http://dx.doi.org/10.1016/j.xpro.2022.101562
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