Nitroxide spin labels and EPR spectroscopy: A powerful association for protein dynamics studies

Site-Directed Spin Labelling (SDSL) technique is based on the attachment of a paramagnetic label onto a specific position of a protein (or other bio-molecules) and the subsequent study by Electron Paramagnetic Resonance (EPR) spectroscopy. In particular, continuous-wave EPR (cw-EPR) spectra can detect the local conformational dynamics for proteins under various conditions. Moreover, pulse-EPR experiments on doubly spin-labelled proteins allow measuring distances between spin centres in the 1.5–8 nm range, providing information about structures and functions. This review focuses on SDSL-EPR spectroscopy as a structural biology tool to investigate proteins using nitroxide labels. The versatility of this spectroscopic approach for protein structural characterization has been demonstrated through the choice of recent studies. The main aim is to provide a general overview of the technique, particularly for non-experts, to spread the applicability of this technique in various fields of structural biology. •SDSL-EPR spectroscopy can provide detailed information about protein structural dynamics.•Nitroxide spin labels are valid tools for the investigation of protein conformations.•Pulse EPR experiments allows to measure distances between nitroxide spin labels in proteins.•Nitroxide spin label report structural information of proteins inside cells.