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  • 1Department of Chemical Physics, Weizmann Institute of Science, Rehovot 7610001, Israel. Daniella.Goldfarb@weizmann.ac.il.

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Relaxation-Induced Dipolar Modulation Enhancement (RIDME) offers superior sensitivity over Double Electron-Electron Resonance (DEER) for measuring distances between Gadolinium (Gd(iii)) spin labels in complex biological systems.

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Area of Science:

  • Electron Paramagnetic Resonance (EPR) Spectroscopy
  • Biophysics
  • Materials Science

Background:

  • Pulse electron paramagnetic resonance methods enable measurement of electron-electron dipolar coupling between spin labels.
  • Gadolinium(iii) (Gd(iii)) spin labels with small zero-field splitting (ZFS) offer high EPR sensitivity due to their narrow central transition.
  • Accurate distance measurements are crucial for understanding molecular structure and dynamics.

Purpose of the Study:

  • To compare the performance of Relaxation-Induced Dipolar Modulation Enhancement (RIDME) and Double Electron-Electron Resonance (DEER/PELDOR) for Gd(iii)-Gd(iii) distance measurements.
  • To evaluate these techniques using both a rigid model compound and protein systems with varying inter-spin distances and distributions.
  • To assess the sensitivity and limitations of RIDME, particularly at W-band frequencies.

Main Methods:

  • Utilized W-band (94.9 GHz, ≈3.5 T) pulse EPR spectroscopy.
  • Employed the dead-time free 5-pulse RIDME sequence and the DEER/PELDOR technique.
  • Investigated a rigid model compound (2.35 nm distance) and two homodimeric protein mutants labeled with DOTA-based Gd(iii) chelates (≈6 nm distance, varying distributions).

Main Results:

  • RIDME demonstrated reduced sensitivity to the failure of the weak coupling approximation compared to DEER for short Gd(iii)-Gd(iii) distances.
  • For longer distances in protein systems, RIDME showed complications from harmonics of the dipolar interaction frequency and increased background subtraction uncertainties.
  • RIDME consistently exhibited significantly higher sensitivity than DEER for both investigated systems.

Conclusions:

  • RIDME is a highly sensitive technique for Gd(iii)-Gd(iii) distance measurements, outperforming DEER, especially for systems with narrow central transitions.
  • While RIDME has specific challenges for S > 1/2 spin systems and background subtraction, its superior sensitivity is advantageous.
  • Further investigation into experimental parameter effects on RIDME traces is warranted for optimizing its application in complex systems.