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

J Biomol NMR. 2011 Dec;51(4):477-85. doi: 10.1007/s10858-011-9578-1. Epub 2011 Oct 25.

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(DQα) shifts and of a substantial fraction of (2)H(DQβ) shifts were obtained for ubiquitin. In the case of OmpG, (2)H(DQα) and (2)H(DQβ) 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Outer Membrane Proteins / chemistry*
  • Bacterial Outer Membrane Proteins / metabolism
  • Carbon / chemistry
  • Carbon / metabolism
  • Deuterium / chemistry
  • Deuterium / metabolism*
  • Escherichia coli / cytology
  • Escherichia coli / metabolism
  • Escherichia coli Proteins / chemistry*
  • Escherichia coli Proteins / metabolism
  • Humans
  • Hydrogen / chemistry
  • Hydrogen / metabolism
  • Isotope Labeling / methods
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Porins / chemistry*
  • Porins / metabolism
  • Proteins / chemistry*
  • Proteins / metabolism
  • Ubiquitin / chemistry*
  • Ubiquitin / metabolism

Substances

  • Bacterial Outer Membrane Proteins
  • Escherichia coli Proteins
  • OmpG protein, E coli
  • Porins
  • Proteins
  • Ubiquitin
  • Carbon
  • Hydrogen
  • Deuterium