Quadruple‐Resonance Magic‐Angle Spinning NMR Spectroscopy of Deuterated Solid Proteins1. (29th January 2014)

• Record Type: Journal Article
• Authors: Akbey, Ümit; Nieuwkoop, Andrew J.; Wegner, Sebastian; Voreck, Anja; Kunert, Britta; Bandara, Priyanga; Engelke, Frank; Nielsen, Niels Chr.; Oschkinat, Hartmut
• Is Part Of: Angewandte Chemie. Volume 126:Number 9(2014)
• Page Start: 2470
• Page End: 2474
• Publication Date: 2014-01-29
• Subjects: Chemistry -- Periodicals; 540
• DOI: 10.1002/ange.201308927
• Languages: English
• ISSNs: 0044-8249

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>Abstract</title> <p> ¹H‐detected magic‐angle spinning NMR experiments facilitate structural biology of solid proteins, which requires using deuterated proteins. However, often amide protons cannot be back‐exchanged sufficiently, because of a possible lack of solvent exposure. For such systems, using ²H excitation instead of ¹H excitation can be beneficial because of the larger abundance and shorter longitudinal relaxation time, <italic>T₁</italic>, of deuterium. A new structure determination approach, "quadruple‐resonance NMR spectroscopy", is presented which relies on an efficient ²H‐excitation and ²H‐¹³C cross‐polarization (CP) step, combined with ¹H detection. We show that by using ²H‐excited experiments better sensitivity is possible on an SH3 sample recrystallized from 30 % H₂O. For a membrane protein, the ABC transporter ArtMP in native lipid bilayers, different sets of signals can be observed from different initial polarization pathways, which can be evaluated further to extract structural properties.</p> </abstract>