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  • Title: Solid-state NMR spectroscopy of oriented membrane polypeptides at 100 K with signal enhancement by dynamic nuclear polarization.
    Author: Salnikov E, Rosay M, Pawsey S, Ouari O, Tordo P, Bechinger B.
    Journal: J Am Chem Soc; 2010 May 05; 132(17):5940-1. PubMed ID: 20392100.
    Abstract:
    Oriented membrane samples encompassing the biradical bTbK and a transmembrane peptide carrying a single (15)N labeled residue have been prepared on polymer sheets with sample geometries that fit into a 3.2 mm MAS rotor. The proton-decoupled (15)N cross-polarization spectra of the peptide were characterized by a single line at fast magic angle spinning speeds of approximately 8 kHz. Irradiating these samples with mu-waves resulted in Dynamic Nuclear Polarization and a concomitant 18-fold signal enhancement which considerably shortened the NMR acquisition times. Furthermore, the sideband patterns of magic angle oriented sample spinning (MAOSS) solid-state NMR spectra at slow spinning speeds (approximately 1 kHz) are indicative that the lipids and peptides form well-oriented bilayers at 100 K despite the narrow inner diameter of the rotor (2.2 mm) and the presence of considerable amounts of biradicals. The DNP signal enhancement opens up enhanced possibilities for multidimensional solid-state NMR investigation of oriented membrane polypeptides.
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