HDX-guided EPR spectroscopy to interrogate membrane protein dynamics

Benjamin J. Lane; Bolin Wang; Yue Ma; Antonio N. Calabrese; Hassane El Mkami; Christos Pliotas

doi:10.1016/j.xpro.2022.101562

HDX-guided EPR spectroscopy to interrogate membrane protein dynamics

Benjamin J. Lane, Bolin Wang, Yue Ma, Antonio N. Calabrese, Hassane El Mkami, Christos Pliotas

Research output: Contribution to journal › Article › peer-review

Abstract

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.

Original language	English
Article number	101562
Number of pages	28
Journal	STAR protocols
Volume	3
Issue number	3
Early online date	18 Jul 2022
DOIs	https://doi.org/10.1016/j.xpro.2022.101562
Publication status	Published - 16 Sept 2022

Bibliographical note

Copyright © The Author(s). This is an open access article distributed under the terms of the Creative Commons CC-BY (https://creativecommons.org/licenses/by/4.0/) license, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Data Access Statement

Data are available within the following link: http://archive.researchdata.leeds.ac.uk/777/ and via the https://doi.org/10.5518/914.
This paper does not report original code. Any additional information required to reanalyze the data reported is available from the lead contact upon request.

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10.1016/j.xpro.2022.101562Licence: CC BY 4.0

HDX-guided EPR spectroscopy to interrogate membrane protein dynamics
Copyright © The Author(s). This is an open access article distributed under the terms of the Creative Commons CC-BY (https://creativecommons.org/licenses/by/4.0/) license, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Final published version, 3.46 MBLicence: CC BY 4.0

Cite this

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title = "HDX-guided EPR spectroscopy to interrogate membrane protein dynamics",

abstract = "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.",

author = "Lane, {Benjamin J.} and Bolin Wang and Yue Ma and Calabrese, {Antonio N.} and {El Mkami}, Hassane and Christos Pliotas",

note = "Copyright {\textcopyright} The Author(s). This is an open access article distributed under the terms of the Creative Commons CC-BY (https://creativecommons.org/licenses/by/4.0/) license, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.",

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T1 - HDX-guided EPR spectroscopy to interrogate membrane protein dynamics

AU - Lane, Benjamin J.

AU - Wang, Bolin

AU - Ma, Yue

AU - Calabrese, Antonio N.

AU - El Mkami, Hassane

AU - Pliotas, Christos

N1 - Copyright © The Author(s). This is an open access article distributed under the terms of the Creative Commons CC-BY (https://creativecommons.org/licenses/by/4.0/) license, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

PY - 2022/9/16

Y1 - 2022/9/16

N2 - 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.

AB - 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.

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JO - STAR protocols

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HDX-guided EPR spectroscopy to interrogate membrane protein dynamics

Abstract

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Data Access Statement

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