Rapid Analysis of DEER Signals Including Short Distances.

Double electron electron resonance (DEER) spectroscopy is an important technique to measure distance distributions P(r) for studying protein structures and protein-protein interactions. DEER data analysis can at times become challenging due to the lack of a detailed analytical signal expression or numerical methods with rapid computation time. We have derived an analytical expression κFULL, which includes both the pseudo-secular dipolar coupling (PSDC) and the finite pulse effects, especially important for shorter distances. Analyses of experiments by κFULL yield accurate and consistent P(r) values for three DEER nitroxide-rulers with distances (rAVG) in the range of 15 to 32 Å, while the current standard analysis produces erroneous results for rAVG < 20 Å. Computation times for deriving P(r) vary between 1 min and 4 min, which is usually much shorter than previous methods that include pseudo-secular and other effects. The expression can be applied to all types of DEER spin probes with little or no modifications.