Quadruple-resonance magic-angle spinning NMR spectroscopy of deuterated solid proteins

Ümit Akbey, Andrew J. Nieuwkoop, Sebastian Wegner, Anja Voreck, Britta Kunert, Priyanga Bandara, Frank Engelke, Niels Chr Nielsen, Hartmut Oschkinat

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

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, T1, 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. Proton-detected magic-angle spinning solid-state NMR spectroscopy using deuterated proteins facilitates structural biology. Solid deuterated proteins that cannot be unfolded/refolded to exchange the deuterons back with protons and proteins that can only be studied in their native environments show a low intrinsic sensitivity in NMR experiments. For such systems, initial excitation of the deuterons can be very useful.

Original languageEnglish (US)
Pages (from-to)2438-2442
Number of pages5
JournalAngewandte Chemie - International Edition
Volume53
Issue number9
DOIs
StatePublished - Feb 24 2014
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Catalysis

Keywords

  • deuteration
  • high sensitivity
  • proteins
  • quadruple-resonance MAS NMR spectroscopy
  • structure elucidation

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