Three-dimensional deuterium-carbon correlation experiments for high-resolution solid-state MAS NMR spectroscopy of large proteins

Daniela Lalli, Paul Schanda, Anup Chowdhury, Joren Retel, Matthias Hiller, Victoria A. Higman, Lieselotte Handel, Vipin Agarwal, Bernd Reif, Barth van Rossum, Umit Akbey, Hartmut Oschkinat*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

17 Citations (Scopus)

Abstract

Well-resolved (2)H-(13)C correlation spectra, reminiscent of (1)H-(13)C correlations, are obtained for perdeuterated ubiquitin and for perdeuterated outer-membrane protein G (OmpG) from E. coli by exploiting the favorable lifetime of (2)H double-quantum (DQ) states. Sufficient signal-to-noise was achieved due to the short deuterium T (1), allowing for high repetition rates and enabling 3D experiments with a (2)H-(13)C transfer step in a reasonable time. Well-resolved 3D (2)H(DQ)-(13)C-(13)C correlations of ubiquitin and OmpG were recorded within 3.5 days each. An essentially complete assignment of (2)H(DQ alpha) shifts and of a substantial fraction of (2)H(DQ beta) shifts were obtained for ubiquitin. In the case of OmpG, (2)H(DQ alpha) and (2)H(DQ beta) chemical shifts of a considerable number of threonine, serine and leucine residues were assigned. This approach provides the basis for a general heteronuclear 3D MAS NMR assignment concept utilizing pulse sequences with (2)H(DQ)-(13)C transfer steps and evolution of deuterium double-quantum chemical shifts.

Original languageEnglish
Pages (from-to)477-485
Number of pages9
JournalJournal of Biomolecular NMR
Volume51
Issue number4
DOIs
Publication statusPublished - Dec 2011

Keywords

  • TRIPLE-RESONANCE NMR
  • Deuterium-carbon correlations
  • ROTATING SOLIDS
  • MAGNETIZATION
  • OmpG
  • DYNAMICS
  • Membrane proteins
  • DOUBLE-QUANTUM NMR
  • PERDEUTERATED PROTEINS
  • SPECTRA
  • ASSIGNMENT
  • ANGLE-SPINNING NMR
  • CHEMICAL-SHIFTS
  • Micro-crystalline
  • Solid-state NMR
  • Ubiquitin

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