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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 L. | |
dc.contributor.author | Ackermann, Katrin | |
dc.contributor.author | Giannoulis, Angeliki | |
dc.contributor.author | Stewart, Alan J. | |
dc.contributor.author | Norman, David G. | |
dc.contributor.author | Bode, Bela E. | |
dc.date.accessioned | 2019-08-16T16:30:03Z | |
dc.date.available | 2019-08-16T16:30:03Z | |
dc.date.issued | 2019-08-19 | |
dc.identifier | 260618369 | |
dc.identifier | 2774d1b0-fe97-4d6a-8092-66840d165160 | |
dc.identifier | 85069937480 | |
dc.identifier | 000480287500018 | |
dc.identifier.citation | Wort , J L , Ackermann , K , Giannoulis , A , Stewart , A J , Norman , D G & Bode , B E 2019 , ' Sub-micromolar pulse dipolar EPR spectroscopy reveals increasing Cu II -labelling of double-histidine motifs with lower temperature ' , Angewandte Chemie - International Edition , vol. 58 , no. 34 , pp. 11681-11685 . https://doi.org/10.1002/anie.201904848 | en |
dc.identifier.issn | 1433-7851 | |
dc.identifier.other | ORCID: /0000-0003-4580-1840/work/60630673 | |
dc.identifier.other | ORCID: /0000-0002-3384-271X/work/60630825 | |
dc.identifier.uri | https://hdl.handle.net/10023/18326 | |
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 | 5 | |
dc.format.extent | 1497312 | |
dc.language.iso | eng | |
dc.relation.ispartof | Angewandte Chemie - International Edition | en |
dc.subject | Dissociation constant | en |
dc.subject | Double-histidine motif | en |
dc.subject | EPR spectroscopy | en |
dc.subject | Non-covalent interactions | en |
dc.subject | RIDME | en |
dc.subject | QD Chemistry | en |
dc.subject | Catalysis | en |
dc.subject | Chemistry(all) | en |
dc.subject | NDAS | 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. School of Medicine | en |
dc.contributor.institution | University of St Andrews. Institute of Behavioural and Neural Sciences | en |
dc.contributor.institution | University of St Andrews. Biomedical Sciences Research Complex | en |
dc.contributor.institution | University of St Andrews. Cellular Medicine Division | en |
dc.contributor.institution | University of St Andrews. Centre of Magnetic Resonance | en |
dc.contributor.institution | University of St Andrews. EaSTCHEM | en |
dc.identifier.doi | 10.1002/anie.201904848 | |
dc.description.status | Peer reviewed | 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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