Lane, Benjamin J., Wang, Bolin, Ma, Yue, Calabrese, Antonio N., El Mkami, Hassane and Pliotas, Christos (2022). HDX-guided EPR spectroscopy to interrogate membrane protein dynamics. STAR protocols, 3 (3),
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.
| Publication DOI: | https://doi.org/10.1016/j.xpro.2022.101562 |
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| Divisions: | College of Health & Life Sciences Aston University (General) |
| Funding Information: | This project was supported by a Biotechnology and Biological Sciences Research Council (BBSRC) grant (BB/S018069/1) to C.P., who also acknowledges support from the Wellcome Trust (WT) (219999/Z/19/Z) and the Chinese Scholarship Council (CSC) in the form o |
| Additional Information: | 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. |
| Publication ISSN: | 2666-1667 |
| 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.<br/>This paper does not report original code. Any additional information required to reanalyze the data reported is available from the lead contact upon request. |
| Last Modified: | 10 Mar 2025 08:24 |
| Date Deposited: | 09 Jan 2025 10:30 |
| Full Text Link: | |
| Related URLs: |
https://www.sci ... 4427?via%3Dihub
(Publisher URL) |
PURE Output Type: | Article |
| Published Date: | 2022-09-16 |
| Published Online Date: | 2022-07-18 |
| Accepted Date: | 2022-07-01 |
| Authors: |
Lane, Benjamin J.
(
0000-0002-3563-2716)
Wang, Bolin Ma, Yue Calabrese, Antonio N. El Mkami, Hassane Pliotas, Christos |