In-Cell Double Electron–Electron Resonance at Nanomolar Protein Concentrations
Svetlana Kucher, Christina Elsner, Mariya Safonova, Stefano Maffini, Enrica Bordignon
Abstract
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.
Topics & Concepts
Electron paramagnetic resonanceSite-directed spin labelingElectron paramagnetic resonance spectroscopyBiomoleculePulsed EPRChemistryGadoliniumElectron nuclear double resonanceNuclear magnetic resonanceResonance (particle physics)BiophysicsSpectroscopyMagnetic resonance imagingSpin echoBiochemistryBiologyPhysicsAtomic physicsMedicineQuantum mechanicsRadiologyOrganic chemistryElectron Spin Resonance StudiesLanthanide and Transition Metal ComplexesAdvanced MRI Techniques and Applications