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HDX-guided EPR spectroscopy to interrogate membrane protein dynamics

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dc.contributor.authorLane, Benjamin J.
dc.contributor.authorWang, Bolin
dc.contributor.authorMa, Yue
dc.contributor.authorCalabrese, Antonio N.
dc.contributor.authorEl Mkami, Hassane
dc.contributor.authorPliotas, Christos
dc.date.accessioned2022-09-14T14:30:07Z
dc.date.available2022-09-14T14:30:07Z
dc.date.issued2022-09-16
dc.identifier280572061
dc.identifier4686b695-949e-43b4-8eb4-44350e47efc9
dc.identifier85134682807
dc.identifier.citationLane , B J , Wang , B , Ma , Y , Calabrese , A N , El Mkami , H & Pliotas , C 2022 , ' HDX-guided EPR spectroscopy to interrogate membrane protein dynamics ' , STAR Protocols , vol. 3 , no. 3 , 101562 . https://doi.org/10.1016/j.xpro.2022.101562en
dc.identifier.issn2666-1667
dc.identifier.otherRIS: urn:EBA98B783C62ADC2D7FD3957562F87BF
dc.identifier.otherPubMedCentral: PMC9304679
dc.identifier.urihttps://hdl.handle.net/10023/26012
dc.descriptionFunding: 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 of studentships for B.J.L. and B.W., respectively. A.N.C. is a Sir Henry Dale Fellow jointly funded by the WT and the Royal Society (220628/Z/20/Z). Funding from the BBSRC (BB/M012573/1) enabled the purchase of mass spectrometry equipment.en
dc.description.abstractSolvent 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).
dc.format.extent28
dc.format.extent3808460
dc.language.isoeng
dc.relation.ispartofSTAR Protocolsen
dc.subjectBiophysicsen
dc.subjectCell membraneen
dc.subjectMolecular biologyen
dc.subjectProtein biochemistryen
dc.subjectProtein expression and purificationen
dc.subjectStructural biologyen
dc.subjectMass spectrometryen
dc.subjectQH301 Biologyen
dc.subjectACen
dc.subjectMCCen
dc.subject.lccQH301en
dc.titleHDX-guided EPR spectroscopy to interrogate membrane protein dynamicsen
dc.typeJournal itemen
dc.contributor.institutionUniversity of St Andrews. School of Physics and Astronomyen
dc.identifier.doi10.1016/j.xpro.2022.101562
dc.description.statusPeer revieweden


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