4.3 Review

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

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ELSEVIER
DOI: 10.1016/j.bbapap.2021.140653

Keywords

SDSL; EPR; DEER; Nitroxide radicals; In-cell EPR

Funding

  1. Regione Toscana
  2. Italian Ministry for University research (FOE funding)
  3. Agence National de la Recherche [ANR-18-CE11-0007-01, ANR-18-0003 NANO-DISPRO]
  4. Conseil Regional Region Sud (AP PhD Fellowship EJD-2018-2021)
  5. Agence Nationale de la Recherche (ANR) [ANR-18-CE11-0007] Funding Source: Agence Nationale de la Recherche (ANR)

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SDSL-EPR technique involves attaching a paramagnetic label to specific positions of proteins for EPR spectroscopic study, providing information on local conformational dynamics, distances between spin centers, structures, and functions. The versatility of this spectroscopic approach in protein structural characterization has been demonstrated through recent studies, aiming to spread its applicability in various fields of structural biology, particularly for non-experts.
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.

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