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dc.contributor.authorWort, Joshua
dc.contributor.authorArya, Swati
dc.contributor.authorAckermann, Katrin
dc.contributor.authorStewart, Alan J.
dc.contributor.authorBode, Bela Ernest
dc.date.accessioned2021-05-06T16:30:16Z
dc.date.available2021-05-06T16:30:16Z
dc.date.issued2021-03-25
dc.identifier.citationWort , J , Arya , S , Ackermann , K , Stewart , A J & Bode , B E 2021 , ' Pulse dipolar EPR reveals double-histidine motif Cu II -NTA spin-labelling robustness against competitor ions ' , Journal of Physical Chemistry Letters , vol. 12 , no. 11 , pp. 2815-2819 . https://doi.org/10.1021/acs.jpclett.1c00211en
dc.identifier.issn1948-7185
dc.identifier.otherPURE: 273246860
dc.identifier.otherPURE UUID: 650e18da-b6aa-40e8-ad34-3b8d5eefa536
dc.identifier.otherORCID: /0000-0003-4580-1840/work/90951871
dc.identifier.otherORCID: /0000-0002-3384-271X/work/90951994
dc.identifier.otherORCID: /0000-0001-7978-9507/work/90952164
dc.identifier.otherScopus: 85103607709
dc.identifier.otherWOS: 000635441500016
dc.identifier.urihttps://hdl.handle.net/10023/23126
dc.descriptionFunding: JLW is supported by the BBSRC DTP Eastbio. We thank the Leverhulme Trust for support (RPG-2018-397). This work was supported by equipment funding through the Wellcome Trust (099149/Z/12/Z) and BBSRC (BB/R013780/1). We gratefully acknowledge ISSF support to the University of St Andrews from the Wellcome Trust.en
dc.description.abstractPulse-dipolar EPR is an appealing strategy for structural characterization of complex systems in solution that complements other biophysical techniques. Significantly, the emergence of genetically encoded self-assembling spin labels exploiting exogenously introduced double-histidine motifs in conjunction with CuII-chelates offers high precision distance determination in systems nonpermissive to thiol-directed spin labeling. However, the noncovalency of this interaction exposes potential vulnerabilities to competition from adventitious divalent metal ions, and pH sensitivity. Herein, a combination of room-temperature isothermal titration calorimetry (ITC) and cryogenic relaxation-induced dipolar modulation enhancement (RIDME) measurements are applied to the model protein Streptococcus sp. group G. protein G, B1 domain (GB1). Results demonstrate double-histidine motif spin labeling using CuII-nitrilotriacetic acid (CuII–NTA) is robust against the competitor ligand ZnII–NTA at >1000-fold molar excess, and high nM binding affinity is surprisingly retained under acidic and basic conditions even though room temperature affinity shows a stronger pH dependence. This indicates the strategy is well-suited for diverse biological applications, with the requirement of other metal ion cofactors or slightly acidic pH not necessarily being prohibitive.
dc.format.extent5
dc.language.isoeng
dc.relation.ispartofJournal of Physical Chemistry Lettersen
dc.rightsCopyright © 2021 The Authors. Published by American Chemical Society. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.en
dc.subjectQC Physicsen
dc.subjectQD Chemistryen
dc.subjectTK Electrical engineering. Electronics Nuclear engineeringen
dc.subjectDASen
dc.subject.lccQCen
dc.subject.lccQDen
dc.subject.lccTKen
dc.titlePulse dipolar EPR reveals double-histidine motif CuII-NTA spin-labelling robustness against competitor ionsen
dc.typeJournal articleen
dc.contributor.sponsorThe Leverhulme Trusten
dc.contributor.sponsorThe Wellcome Trusten
dc.contributor.sponsorBBSRCen
dc.contributor.sponsorThe Leverhulme Trusten
dc.contributor.sponsorThe Wellcome Trusten
dc.description.versionPublisher PDFen
dc.contributor.institutionUniversity of St Andrews. School of Chemistryen
dc.contributor.institutionUniversity of St Andrews. Cellular Medicine Divisionen
dc.contributor.institutionUniversity of St Andrews. School of Medicineen
dc.contributor.institutionUniversity of St Andrews. Sir James Mackenzie Institute for Early Diagnosisen
dc.contributor.institutionUniversity of St Andrews. Biomedical Sciences Research Complexen
dc.contributor.institutionUniversity of St Andrews. Institute of Behavioural and Neural Sciencesen
dc.contributor.institutionUniversity of St Andrews. EaSTCHEMen
dc.contributor.institutionUniversity of St Andrews. Centre of Magnetic Resonanceen
dc.identifier.doihttps://doi.org/10.1021/acs.jpclett.1c00211
dc.description.statusPeer revieweden
dc.identifier.grantnumberRPG-2018-397en
dc.identifier.grantnumber099149/Z/12/Zen
dc.identifier.grantnumberBB/R013780/1en
dc.identifier.grantnumberRPG-2017-214en
dc.identifier.grantnumberen


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