Abstract Well-resolved 2Hâ??13C correlation spectra, reminiscent of 1Hâ??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 2Hâ??13C transfer step in a reasonable time. Well-resolved 3D 2HDQâ??13Câ??13C correlations of ubiquitin and OmpG were recorded within 3.5 days 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 2HDQâ??13C transfer steps and evolution of deuterium double-quantum chemical shifts.
Ultra-high resolution in MAS solid-state NMR of perdeuterated proteins: Implications for Structure and Dynamics
Ultra-high resolution in MAS solid-state NMR of perdeuterated proteins: Implications for Structure and Dynamics
Publication year: 2012
Source: Journal of Magnetic Resonance, Available online 5 January 2012</br>
Bernd*Reif</br>
http://www.sciencedirect.com/cache/MiamiImageURL/1-s2.0-S1090780711005969-fx1.sml</br></br></br>
Source: Journal of Magnetic Resonance
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Authors: Marassi FM, Ma C, Gesell JJ, Opella SJ
Uniformly (15)N-labeled samples of membrane proteins with...
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High Resolution 1H Detected 1H,13C Correlation Spectra in MAS Solid-State NMR using Deuterated Proteins with Selective 1H,2H Isotopic Labeling of Methyl Groups
High Resolution <SUP>1</SUP>H Detected <SUP>1</SUP>H,<SUP>13</SUP>C Correlation Spectra in MAS Solid-State NMR using Deuterated Proteins with Selective <SUP>1</SUP>H,<SUP>2</SUP>H Isotopic Labeling of Methyl Groups
Vipin Agarwal, Anne Diehl, Nikolai Skrynnikov, and Bernd Reif
J. Am. Chem. Soc.; 2006; 128(39) pp 12620 - 12621;
Abstract:
MAS solid-state NMR experiments applied to biological solids are still hampered by low sensitivity and resolution. In this work, we employ a deuteration scheme in which individual methyl groups are selectively protonated. This labeling scheme...
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Four-dimensional heteronuclear correlation experiments for chemical shift assignment of solid proteins
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W. Trent Franks, Kathryn D. Kloepper, Benjamin J. Wylie and Chad M. Rienstra
Journal of Biomolecular NMR; 2007; 39(2); pp 107 - 131
Abstract:
Chemical shift assignment is the first step in all established protocols for structure determination of uniformly labeled proteins by NMR. The explosive growth in recent years of magic-angle spinning (MAS) solid-state NMR (SSNMR) applications is largely attributable to improved methods for backbone and side-chain chemical shift correlation...
Three-dimensional deuterium-carbon correlation experiments for high-resolution solid-state MAS NMR spectroscopy of large proteins
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