title: Biomolecular DNP-Supported NMR Spectroscopy using Site-Directed Spin Labeling.
abstract: Dynamic nuclear polarization (DNP) has been shown to greatly enhance spectroscopic sensitivity, creating novel opportunities for NMR studies on complex and large molecular assemblies in life and material sciences. In such applications, however, site-specificity and spectroscopic resolution become critical factors that are usually difficult to control by current DNP-based approaches. We have examined in detail the effect of directly attaching mono- or biradicals to induce local paramagnetic relaxation effects and, at the same time, to produce sizable DNP enhancements. Using a membrane-embedded ion channel as an example, we varied the degree of paramagnetic labeling and the location of the DNP probes. Our results show that the creation of local spin clusters can generate sizable DNP enhancements while preserving the intrinsic benefits of paramagnetic relaxation enhancement (PRE)-based NMR approaches. DNP using chemical labeling may hence provide an attractive route to introduce molecular specificity into DNP studies in life science applications and beyond.
authors: NMR research group Bijvoet Center for Biomolecular Research, Utrecht University, Padualaan 8, 3584 CH Utrecht (The Netherlands).
authors: Elwin A W
authors: EA
authors: van der Cruijsen
authors: NMR research group Bijvoet Center for Biomolecular Research, Utrecht University, Padualaan 8, 3584 CH Utrecht (The Netherlands).
authors: Eline J
authors: EJ
authors: Koers
authors: Aix Marseille Université, CNRS, ICR UMR 7273, 13397 Marseille (France).
authors: Claire
authors: C
authors: Sauvée
authors: Department of Biochemistry and Molecular Biology, The University of Chicago, 929 E57th Street, Chicago, Illinois 60637 (USA).
authors: Raymond E
authors: RE
authors: Hulse
authors: NMR research group Bijvoet Center for Biomolecular Research, Utrecht University, Padualaan 8, 3584 CH Utrecht (The Netherlands).
authors: Markus
authors: M
authors: Weingarth
authors: Aix Marseille Université, CNRS, ICR UMR 7273, 13397 Marseille (France).
authors: Olivier
authors: O
authors: Ouari
authors: Department of Biochemistry and Molecular Biology, The University of Chicago, 929 E57th Street, Chicago, Illinois 60637 (USA).
authors: Eduardo
authors: E
authors: Perozo
authors: Aix Marseille Université, CNRS, ICR UMR 7273, 13397 Marseille (France).
authors: Paul
authors: P
authors: Tordo
authors: NMR research group Bijvoet Center for Biomolecular Research, Utrecht University, Padualaan 8, 3584 CH Utrecht (The Netherlands). m.baldus@uu.nl.
authors: Marc
authors: M
authors: Baldus
mesh_terms: D004578:Electron Spin Resonance Spectroscopy; D008565:Membrane Proteins / Q000737:chemistry*; D008859:Microscopy, Polarization; D019906:Nuclear Magnetic Resonance, Biomolecular; D013113:Spin Labels
chemical_list: D008565:Membrane Proteins; D013113:Spin Labels
keywords: DNP; KcsA; MTSSL; NMR spectroscopy; membrane proteins; spin labeling