OOP-ESEEM Spectroscopy: Accuracies of Distances of Spin-Correlated Radical Pairs in Biomolecules

In addition to the commonly used electron-electron double resonance (ELDOR) technique, there are several other electron paramagnetic resonance (EPR) methods by which structure information can be obtained by exploiting the dipolar coupling between two radicals based on its characteristic r(-3) dependence. In this contribution, we explore the potential of out-of-phase-electron-spin echo envelope modulation (OOP-ESEEM) spectroscopy to collect accurate distance information in photo-sensitive (bio) molecules. Although the method has already been applied to spin-correlated radical pairs in several classes of light-active proteins, the accuracy of the information obtained has not yet been extensively evaluated. To do this in a system-independent fashion, OOP-ESEEM time traces simulated with different values of the dipolar and exchange couplings were generated and analyzed in a best-possible way. Excellent agreement between calculated and numerically fitted values over a wide range of distances (between 15 and 45 angstrom) was obtained. Furthermore, the limitations of the method and the dependence on various experimental parameters could be evaluated.

Automated summary

This study explores the potential of out-of-phase-electron-spin echo envelope modulation (OOP-ESEEM) spectroscopy in collecting accurate distance information in photo-sensitive (bio) molecules. The OOP-ESEEM time traces were simulated and analyzed to obtain excellent agreement between calculated and numerically fitted values over a wide range of distances.