RIDME distance measurements using Gd(III) tags with a narrow central transition

A. Collauto, Veronica Frydman, M D Lee, E H Abdelkader, A Feintuch, J D Swarbrick, B Graham, G Otting, D Goldfarb

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Methods based on pulse electron paramagnetic resonance allow measurement of the electron-electron dipolar coupling between two spin labels. Here we compare the most popular technique, Double Electron-Electron Resonance (DEER or PELDOR), with the dead-time free 5-pulse Relaxation-Induced Dipolar Modulation Enhancement (RIDME) method for Gd(iii)-Gd(iii) distance measurements at W-band (94.9 GHz, ≈3.5 T) using Gd(iii) tags with a small zero field splitting (ZFS). Such tags are important because of their high EPR sensitivity arising from their narrow central transition. Two systems were investigated: (i) a rigid model compound with an inter-spin distance of 2.35 nm, and (ii) two mutants of a homodimeric protein, both labeled with a DOTA-based Gd(iii) chelate and characterized by an inter-spin distance of around 6 nm, one having a narrow distance distribution and the other a broad distribution. Measurements on the model compound show that RIDME is less sensitive to the complications arising from the failure of the weak coupling approximation which affect DEER measurements on systems characterized by short inter-spin distances between Gd(iii) tags having a narrow central transition. Measurements on the protein samples, which are characterized by a long inter-spin distance, emphasize the complications due to the appearance of harmonics of the dipolar interaction frequency in the RIDME traces for S > 1/2 spin systems, as well as enhanced uncertainties in the background subtraction. In both cases the sensitivity of RIDME was found to be significantly better than DEER. The effects of the experimental parameters on the RIDME trace are discussed.

Original languageEnglish
Pages (from-to)19037-19049
Number of pages13
JournalPhysical Chemistry Chemical Physics
Volume18
Issue number28
DOIs
Publication statusPublished - 2016

Cite this

Collauto, A., Frydman, V., Lee, M. D., Abdelkader, E. H., Feintuch, A., Swarbrick, J. D., ... Goldfarb, D. (2016). RIDME distance measurements using Gd(III) tags with a narrow central transition. Physical Chemistry Chemical Physics, 18(28), 19037-19049. https://doi.org/10.1039/C6CP03299K
Collauto, A. ; Frydman, Veronica ; Lee, M D ; Abdelkader, E H ; Feintuch, A ; Swarbrick, J D ; Graham, B ; Otting, G ; Goldfarb, D. / RIDME distance measurements using Gd(III) tags with a narrow central transition. In: Physical Chemistry Chemical Physics. 2016 ; Vol. 18, No. 28. pp. 19037-19049.
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abstract = "Methods based on pulse electron paramagnetic resonance allow measurement of the electron-electron dipolar coupling between two spin labels. Here we compare the most popular technique, Double Electron-Electron Resonance (DEER or PELDOR), with the dead-time free 5-pulse Relaxation-Induced Dipolar Modulation Enhancement (RIDME) method for Gd(iii)-Gd(iii) distance measurements at W-band (94.9 GHz, ≈3.5 T) using Gd(iii) tags with a small zero field splitting (ZFS). Such tags are important because of their high EPR sensitivity arising from their narrow central transition. Two systems were investigated: (i) a rigid model compound with an inter-spin distance of 2.35 nm, and (ii) two mutants of a homodimeric protein, both labeled with a DOTA-based Gd(iii) chelate and characterized by an inter-spin distance of around 6 nm, one having a narrow distance distribution and the other a broad distribution. Measurements on the model compound show that RIDME is less sensitive to the complications arising from the failure of the weak coupling approximation which affect DEER measurements on systems characterized by short inter-spin distances between Gd(iii) tags having a narrow central transition. Measurements on the protein samples, which are characterized by a long inter-spin distance, emphasize the complications due to the appearance of harmonics of the dipolar interaction frequency in the RIDME traces for S > 1/2 spin systems, as well as enhanced uncertainties in the background subtraction. In both cases the sensitivity of RIDME was found to be significantly better than DEER. The effects of the experimental parameters on the RIDME trace are discussed.",
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Collauto, A, Frydman, V, Lee, MD, Abdelkader, EH, Feintuch, A, Swarbrick, JD, Graham, B, Otting, G & Goldfarb, D 2016, 'RIDME distance measurements using Gd(III) tags with a narrow central transition' Physical Chemistry Chemical Physics, vol. 18, no. 28, pp. 19037-19049. https://doi.org/10.1039/C6CP03299K

RIDME distance measurements using Gd(III) tags with a narrow central transition. / Collauto, A.; Frydman, Veronica; Lee, M D; Abdelkader, E H; Feintuch, A; Swarbrick, J D; Graham, B; Otting, G; Goldfarb, D.

In: Physical Chemistry Chemical Physics, Vol. 18, No. 28, 2016, p. 19037-19049.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Collauto, A.

AU - Frydman, Veronica

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AU - Abdelkader, E H

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AU - Swarbrick, J D

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AB - Methods based on pulse electron paramagnetic resonance allow measurement of the electron-electron dipolar coupling between two spin labels. Here we compare the most popular technique, Double Electron-Electron Resonance (DEER or PELDOR), with the dead-time free 5-pulse Relaxation-Induced Dipolar Modulation Enhancement (RIDME) method for Gd(iii)-Gd(iii) distance measurements at W-band (94.9 GHz, ≈3.5 T) using Gd(iii) tags with a small zero field splitting (ZFS). Such tags are important because of their high EPR sensitivity arising from their narrow central transition. Two systems were investigated: (i) a rigid model compound with an inter-spin distance of 2.35 nm, and (ii) two mutants of a homodimeric protein, both labeled with a DOTA-based Gd(iii) chelate and characterized by an inter-spin distance of around 6 nm, one having a narrow distance distribution and the other a broad distribution. Measurements on the model compound show that RIDME is less sensitive to the complications arising from the failure of the weak coupling approximation which affect DEER measurements on systems characterized by short inter-spin distances between Gd(iii) tags having a narrow central transition. Measurements on the protein samples, which are characterized by a long inter-spin distance, emphasize the complications due to the appearance of harmonics of the dipolar interaction frequency in the RIDME traces for S > 1/2 spin systems, as well as enhanced uncertainties in the background subtraction. In both cases the sensitivity of RIDME was found to be significantly better than DEER. The effects of the experimental parameters on the RIDME trace are discussed.

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Collauto A, Frydman V, Lee MD, Abdelkader EH, Feintuch A, Swarbrick JD et al. RIDME distance measurements using Gd(III) tags with a narrow central transition. Physical Chemistry Chemical Physics. 2016;18(28):19037-19049. https://doi.org/10.1039/C6CP03299K