Lane, BJ orcid.org/0000-0002-3563-2716, Wang, B orcid.org/0000-0003-2801-372X, Ma, Y et al. (3 more authors) (2022) HDX-guided EPR spectroscopy to interrogate membrane protein dynamics. STAR Protocols, 3 (3). 101562. 101562-. ISSN 2666-1667
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.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2022 The Author(s). This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
Keywords: | Biophysics; Cell Membrane; Mass Spectrometry; Molecular Biology; Protein Biochemistry; Protein expression and purification; Structural Biology; Electron Spin Resonance Spectroscopy; Membrane Proteins; Spin Labels |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Biomedical Sciences (Leeds) |
Funding Information: | Funder Grant number BBSRC (Biotechnology & Biological Sciences Research Council) BB/S018069/1 |
Depositing User: | Symplectic Publications |
Date Deposited: | 29 Jul 2022 09:54 |
Last Modified: | 08 Jan 2025 12:49 |
Status: | Published |
Publisher: | Cell Press |
Identification Number: | 10.1016/j.xpro.2022.101562 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:189476 |