Three-dimensional deuterium-carbon correlation experiments for high-resolution solid-state MAS NMR spectroscopy of large proteins
Lalli, Daniela ; Schanda, Paul ; Chowdhury, Anup ; Retel, Joren ; Hiller, Matthias ; Higman, Victoria ; Handel, Lieselotte ; Agarwal, Vipin ; Reif, Bernd ; van Rossum, Barth ; Akbey, Ümit ; Oschkinat, Hartmut
In: Journal of Biomolecular NMR, 2011, vol. 51, no. 4, p. 477-485
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- Well-resolved 2H-13C correlation spectra, reminiscent of 1H-13C correlations, are obtained for perdeuterated ubiquitin and for perdeuterated outer-membrane protein G (OmpG) from E. coli by exploiting the favorable lifetime of 2H 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 2H-13C transfer step in a reasonable time. Well-resolved 3D 2HDQ-13C-13C correlations of ubiquitin and OmpG were recorded within 3.5days each. An essentially complete assignment of 2HDQα shifts and of a substantial fraction of 2HDQβ shifts were obtained for ubiquitin. In the case of OmpG, 2HDQα and 2HDQβ 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 2HDQ-13C transfer steps and evolution of deuterium double-quantum chemical shifts