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Asami, S.* ; Schmieder, P.* ; Reif, B.

High resolution 1H-detected solid-state NMR spectroscopy of protein aliphatic resonances: Access to tertiary structure information.

J. Am. Chem. Soc. 132, 15133-15135 (2010)
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Biological magic angle spinning (MAS) solid-state nuclear magnetic resonance spectroscopy has developed rapidly over the past two decades. For the structure determination of a protein by solid-state NMR, routinely C-13,C-13 distance restraints as well as dihedral restraints are employed. In protonated samples, this is achieved by growing the bacterium on a medium which contains [1,3]-C-13 glycerol or [2]-C-13 glycerol to dilute the C-13 spin system. Labeling schemes, which rely on heteronuclei, are insensitive both for detection and in terms of quantification of distances, since they are relying on low-gamma nuclei. Proton detection can in principle provide a gain in sensitivity by a factor of 8 and 31, compared to the C-13 or N-15 detected version of the experiment. We report here a new labeling scheme, which enables H-1-detection of aliphatic resonances with high resolution in MAS solid-state NMR spectroscopy. We prepared microcrystals of the SH3 domain of chicken a-spectrin with 5% protonation at nonexchangeable sites and obtained line widths on the order of 25 Hz for aliphatic H-1 resonances. We show further that C-13 resolved 3D-H-1,H-1 correlation experiments yield access to long-range proton-proton distances in the protein.
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Publication type Article: Journal article
Document type Scientific Article
Keywords PERDEUTERATED PROTEINS; ROTATING SOLIDS; SH3 DOMAIN; CONSTRAINTS; DEUTERATION; PEPTIDES
ISSN (print) / ISBN 0002-7863
e-ISSN 1520-5126
Quellenangaben Volume: 132, Issue: 43, Pages: 15133-15135 Article Number: , Supplement: ,
Publisher American Chemical Society (ACS)
Publishing Place Washington
Reviewing status Peer reviewed