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Measuring Nanometer Distances in Proteins and Rigid Rulers between 19F and Gd3+ by Integration of 19F-ENDOR Signal
Martyna Judd1, Mian Qi2, Elwy H Abdelkader3
1Research School of Chemistry, Australian National University, Canberra, Australian Capital Territory 2601, Australia.
Journal of the American Chemical Society
|May 6, 2025
Summary
Fluorine-19 ENDOR (Electron Nuclear Double Resonance) now measures longer protein distances up to 20 Å. This technique enhances structural biology by enabling nanometer-scale triangulation of ligand positions in protein complexes.
Area of Science:
- Structural Biology
- Biophysics
- Magnetic Resonance Spectroscopy
Background:
- Fluorine-19 ENDOR (Electron Nuclear Double Resonance) is a valuable technique for measuring distances in proteins.
- Current limitations in spin-spin relaxation and line width restrict inter-tag distance measurements to approximately 15 Å.
Purpose of the Study:
- To extend the measurable distance range of Fluorine-19 ENDOR.
- To develop a robust method for determining longer inter-tag distances in protein structures.
Main Methods:
- Utilized geometrically well-defined protein rulers labeled with Fluorine-19 and a paramagnetic tag (Gadolinium 3+).
- Integrated the intensity of the ENDOR spectrum to determine distances.
- Scaled spectral intensities to a known reference for robustness.
Main Results:
- Demonstrated the ability to measure 19F-Gd3+ distances up to 20 Å.
- Indicated potential for measuring distances approaching 30 Å.
- Established the method's robustness through intensity scaling.
Conclusions:
- Extended the utility of 19F ENDOR for measuring longer distances in structural biology.
- The developed method allows for nanometer-scale triangulation of ligand coordinates within protein-ligand complexes.
