Three-dimensional deuterium-carbon correlation experiments for high-resolution solid-state MAS NMR spectroscopy of large proteins.
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.
Leibniz-Institut für Molekulare Pharmakologie (FMP), Berlin, Germany.
Daniela
D
Lalli
Paul
P
Schanda
Anup
A
Chowdhury
Joren
J
Retel
Matthias
M
Hiller
Victoria A
VA
Higman
Lieselotte
L
Handel
Vipin
V
Agarwal
Bernd
B
Reif
Barth
B
van Rossum
Umit
U
Akbey
Hartmut
H
Oschkinat
D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D002244:Carbon / Q000737:chemistry / Q000378:metabolism; D003903:Deuterium / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000166:cytology / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D006859:Hydrogen / Q000737:chemistry / Q000378:metabolism; D007553:Isotope Labeling / Q000379:methods; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D018272:Porins / Q000737:chemistry* / Q000378:metabolism; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D025801:Ubiquitin / Q000737:chemistry* / Q000378:metabolism
D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; C114347:OmpG protein, E coli; D018272:Porins; D011506:Proteins; D025801:Ubiquitin; D002244:Carbon; D006859:Hydrogen; D003903:Deuterium