Projects per year
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.
Original language | English |
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Pages (from-to) | 11681-11685 |
Number of pages | 5 |
Journal | Angewandte Chemie - International Edition |
Volume | 58 |
Issue number | 34 |
Early online date | 18 Jul 2019 |
DOIs | |
Publication status | Published - 19 Aug 2019 |
Keywords
- Dissociation constant
- Double-histidine motif
- EPR spectroscopy
- Non-covalent interactions
- RIDME
Fingerprint
Dive into the research topics of 'Sub-micromolar pulse dipolar EPR spectroscopy reveals increasing CuII-labelling of double-histidine motifs with lower temperature'. Together they form a unique fingerprint.Projects
- 3 Finished
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Cryogen-free arbitrary waveform EPR: Cryogen-free arbitrary waveform EPR for structural Biology and Biophysics
Bode, B. E. (PI), Lovett, J. E. (CoI), Pliotas, C. (CoI), Schwarz-Linek, U. (CoI), Smith, G. M. (CoI), Stewart, A. J. (CoI) & White, M. (CoI)
1/05/18 → 30/04/19
Project: Standard
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ISSF3 Wellcome Trust Award: Institutional Strategic Support Fund (ISSF3)
Brown, C. T. A. (PI)
1/03/17 → 29/02/20
Project: Standard
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State of the art pulse EPR instrumentati: State of the art pulse EPR instrumentation for long range distance measurements in biomacromolecules
Smith, G. M. (PI), Bode, B. E. (CoI), Naismith, J. (CoI), Schiemann, O. (CoI) & White, M. (CoI)
1/09/12 → 31/08/17
Project: Standard
Datasets
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Data underpinning: Sub-micromolar pulse dipolar EPR spectroscopy reveals increasing Cu(II)-labelling of double-histidine motifs with lower temperature
Wort, J. (Creator), Ackermann, K. (Creator), Giannoulis, A. (Creator), Stewart, A. J. (Creator), Norman, D. (Creator) & Bode, B. E. (Creator), University of St Andrews, 4 Jul 2019
DOI: 10.17630/ced80b64-fa40-4d15-8f17-78bb4f06562f
Dataset
File
Research output
- 1 Article
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Sub-micromolar pulse dipolar EPR spectroscopy reveals increasing CuII-labelling of double-histidine motifs with lower temperature
Wort, J., Ackermann, K., Giannoulis, A., Stewart, A. J., Norman, D. & Bode, B. E., 19 Aug 2019, In: Angewandte Chemie. 131, 34, p. 11807-11811 6 p.Research output: Contribution to journal › Article › peer-review
Open AccessFile