Short-range ENDOR distance measurements between Gd(III) and trifluoromethyl labels in proteins
Judd M, Abdelkader EH, Qi M, Harmer JR, Huber T, Godt A, Savitsky A, Otting G, Cox N (2022)
Physical chemistry chemical physics : PCCP.
Zeitschriftenaufsatz
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Autor*in
Judd, Martyna;
Abdelkader, Elwy H;
Qi, MianUniBi
;
Harmer, Jeffrey R;
Huber, Thomas;
Godt, AdelheidUniBi;
Savitsky, Anton;
Otting, Gottfried;
Cox, Nicholas
![](https://pub.uni-bielefeld.de/images/icon_orcid.png)
Abstract / Bemerkung
The measurement of distances in proteins can be challenging in the 5-20 A range, which is outside those accessible through conventional NMR and EPR methods. Recently it was demonstrated that distances in this range could be measured between a nitroxide as a paramagnetic spin label and a nearby fluorine atom (19F) as a nuclear spin label using high-field (W-band/3.4 T) ENDOR spectroscopy. Here we show that such measurements can also be performed using a gadolinium ion (Gd3+) as the paramagnetic tag. Gd3+ has two advantages. (i) A greater electronic spin (S = 7/2) and fast electronic spin-lattice (T1) relaxation, improving sensitivity by allowing data to be collected at lower temperatures. (ii) A narrow EPR signal for the - transition, and therefore no orientation selection artefacts. Signal intensities can be further enhanced by using a trifluoromethyl (C19F3) group instead of a single 19F atom. Using the protein calbindin D9k with a Ca2+ ion replaced by a Gd3+ ion and a trifluoromethylphenylalanine in position 50, we show that distances up to about 10 A can be readily measured. Longer distances proved more difficult to measure due to variable electronic TM relaxation rates, which lead to broader Lorentzian ENDOR lineshapes. Gd3+ complexes (Gd3+ tags), which reliably display longer TM times, allow longer distances to be measured (8-16 A). We also provide preliminary evidence that the intensity of ENDOR signals follows the predicted 1/r6 dependence, indicating that distances r > 20 A can be measured by this method.
Erscheinungsjahr
2022
Zeitschriftentitel
Physical chemistry chemical physics : PCCP
eISSN
1463-9084
Page URI
https://pub.uni-bielefeld.de/record/2966464
Zitieren
Judd M, Abdelkader EH, Qi M, et al. Short-range ENDOR distance measurements between Gd(III) and trifluoromethyl labels in proteins. Physical chemistry chemical physics : PCCP. 2022.
Judd, M., Abdelkader, E. H., Qi, M., Harmer, J. R., Huber, T., Godt, A., Savitsky, A., et al. (2022). Short-range ENDOR distance measurements between Gd(III) and trifluoromethyl labels in proteins. Physical chemistry chemical physics : PCCP. https://doi.org/10.1039/d2cp02889a
Judd, Martyna, Abdelkader, Elwy H, Qi, Mian, Harmer, Jeffrey R, Huber, Thomas, Godt, Adelheid, Savitsky, Anton, Otting, Gottfried, and Cox, Nicholas. 2022. “Short-range ENDOR distance measurements between Gd(III) and trifluoromethyl labels in proteins”. Physical chemistry chemical physics : PCCP.
Judd, M., Abdelkader, E. H., Qi, M., Harmer, J. R., Huber, T., Godt, A., Savitsky, A., Otting, G., and Cox, N. (2022). Short-range ENDOR distance measurements between Gd(III) and trifluoromethyl labels in proteins. Physical chemistry chemical physics : PCCP.
Judd, M., et al., 2022. Short-range ENDOR distance measurements between Gd(III) and trifluoromethyl labels in proteins. Physical chemistry chemical physics : PCCP.
M. Judd, et al., “Short-range ENDOR distance measurements between Gd(III) and trifluoromethyl labels in proteins”, Physical chemistry chemical physics : PCCP, 2022.
Judd, M., Abdelkader, E.H., Qi, M., Harmer, J.R., Huber, T., Godt, A., Savitsky, A., Otting, G., Cox, N.: Short-range ENDOR distance measurements between Gd(III) and trifluoromethyl labels in proteins. Physical chemistry chemical physics : PCCP. (2022).
Judd, Martyna, Abdelkader, Elwy H, Qi, Mian, Harmer, Jeffrey R, Huber, Thomas, Godt, Adelheid, Savitsky, Anton, Otting, Gottfried, and Cox, Nicholas. “Short-range ENDOR distance measurements between Gd(III) and trifluoromethyl labels in proteins”. Physical chemistry chemical physics : PCCP (2022).
Daten bereitgestellt von European Bioinformatics Institute (EBI)
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