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Sub-micromolar pulse dipolar EPR spectroscopy reveals increasing CuII-labelling of double-histidine motifs with lower temperature
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dc.contributor.author | Wort, Joshua | |
dc.contributor.author | Ackermann, Katrin | |
dc.contributor.author | Giannoulis, Angeliki | |
dc.contributor.author | Stewart, Alan J. | |
dc.contributor.author | Norman, David | |
dc.contributor.author | Bode, Bela E. | |
dc.date.accessioned | 2019-07-19T09:30:05Z | |
dc.date.available | 2019-07-19T09:30:05Z | |
dc.date.issued | 2019-08-19 | |
dc.identifier | 259356601 | |
dc.identifier | d03b8519-5574-4b98-9bad-b615f1c45940 | |
dc.identifier.citation | Wort , J , Ackermann , K , Giannoulis , A , Stewart , A J , Norman , D & Bode , B E 2019 , ' Sub-micromolar pulse dipolar EPR spectroscopy reveals increasing Cu II -labelling of double-histidine motifs with lower temperature ' , Angewandte Chemie , vol. 131 , no. 34 , pp. 11807-11811 . https://doi.org/10.1002/ange.201904848 | en |
dc.identifier.issn | 0044-8249 | |
dc.identifier.other | ORCID: /0000-0002-3384-271X/work/59698756 | |
dc.identifier.other | ORCID: /0000-0003-4580-1840/work/60195777 | |
dc.identifier.uri | https://hdl.handle.net/10023/18131 | |
dc.description | JLW is supported by the BBSRC DTP Eastbio. 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.abstract | Electron paramagnetic resonance (EPR) distance measurements are making increasingly important contributions to the studies of biomolecules by providing highly accurate geometric constraints. Combining double‐histidine motifs with CuII spin labels can further increase the precision of distance measurements. It is also useful for proteins containing essential cysteines that can interfere with thiol‐specific labelling. However, the non‐covalent CuII coordination approach is vulnerable to low binding‐affinity. Herein, dissociation constants (KD) are investigated directly from the modulation depths of relaxation‐induced dipolar modulation enhancement (RIDME) EPR experiments. This reveals low‐ to sub‐μm CuII KDs under EPR distance measurement conditions at cryogenic temperatures. We show the feasibility of exploiting the double‐histidine motif for EPR applications even at sub‐μm protein concentrations in orthogonally labelled CuII–nitroxide systems using a commercial Q‐band EPR instrument. | |
dc.format.extent | 6 | |
dc.format.extent | 1112821 | |
dc.language.iso | eng | |
dc.relation.ispartof | Angewandte Chemie | en |
dc.subject | RIDME | en |
dc.subject | Dissociation-constant | en |
dc.subject | Double-histidine motif | en |
dc.subject | Non-covalent interactions | en |
dc.subject | EPR-spectroscopy | en |
dc.subject | QD Chemistry | en |
dc.subject | DAS | en |
dc.subject | BDC | en |
dc.subject | R2C | en |
dc.subject.lcc | QD | en |
dc.title | Sub-micromolar pulse dipolar EPR spectroscopy reveals increasing CuII-labelling of double-histidine motifs with lower temperature | en |
dc.type | Journal article | en |
dc.contributor.sponsor | BBSRC | en |
dc.contributor.sponsor | The Wellcome Trust | en |
dc.contributor.sponsor | The Wellcome Trust | en |
dc.contributor.institution | University of St Andrews. School of Chemistry | en |
dc.contributor.institution | University of St Andrews. Sir James Mackenzie Institute for Early Diagnosis | en |
dc.contributor.institution | University of St Andrews. Cellular Medicine Division | en |
dc.contributor.institution | University of St Andrews. Biomedical Sciences Research Complex | en |
dc.contributor.institution | University of St Andrews. Institute of Behavioural and Neural Sciences | en |
dc.contributor.institution | University of St Andrews. School of Medicine | en |
dc.contributor.institution | University of St Andrews. EaSTCHEM | en |
dc.contributor.institution | University of St Andrews. Centre of Magnetic Resonance | en |
dc.identifier.doi | https://doi.org/10.1002/ange.201904848 | |
dc.description.status | Peer reviewed | en |
dc.identifier.url | https://onlinelibrary.wiley.com/doi/abs/10.1002/anie.201904848?af=R | en |
dc.identifier.grantnumber | BB/R013780/1 | en |
dc.identifier.grantnumber | 099149/Z/12/Z | en |
dc.identifier.grantnumber | 204821/Z/16/Z | en |
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