In-Cell Double Electron-Electron Resonance at Nanomolar Protein Concentrations

Electron paramagnetic resonance (EPR) spectroscopy is an established technique to site-specifically monitor conformational changes of spin-labeled biomolecules. Emerging in-cell EPR approaches aiming to address spin-labeled proteins in their native environment still struggle to reach a broad applicability and to target physiologically relevant protein concentrations. Here, we present a comparative in vitro and in-cell double electron-electron resonance (DEER) study demonstrating that nanomolar protein concentrations are at reach to measure distances up to 4.5 nm between protein sites carrying commercial gadolinium spin labels.

Automated summary

The study demonstrates the feasibility of using double electron-electron resonance (DEER) technique to measure distances between protein sites at very low protein concentrations, providing a new approach for studying conformational changes of biomolecules.