| Three-dimensional triple-resonance NMR Spectroscopy of isotopically enriched proteins. | |
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MedLine Citation:
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PMID: 22152361 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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Four new and complementary three-dimensional triple-resonance experiments are described for obtaining complete backbone (1)H, (13)C, and (15)N resonance assignments of proteins uniformly enriched with (13)C and (15)N. The new methods all rely on (1)H detection and use multiple magnetization transfers through well-resolved one-bond J couplings. Therefore, the 3D experiments are sensitive and permit relatively rapid recording of 3D spectra (l-2days) for protein concentrations on the order of 1mM. One experiment (HNCO) correlates the amide (1)H and (15)N shifts with the (13)C shift of the carbonyl resonance of the preceding amino acid. A second experiment (HNCA) correlates the intraresidue amide (1)H and (15)N shifts with the Cα chemical shift. This experiment often also provides a weak correlation between the amide NH and (15)N resonances of one amino acid and the Ca resonance of the preceding amino acid. A third experiment (HCACO) correlates the Hα and Cα shifts with the intraresidue carbonyl shift. Finally, a 3D relay experiment, HCA(CO)N, correlates Ha and Cal resonances of one residue with the (15)N frequency of the succeeding residue. The principles of these experiments are described in terms of the operator formalism. To optimize spectral resolution, special attention is paid to removal of undesired J splittings in the 3D spectra. Technical details regarding the implementation of these triple-resonance experiments on a commercial spectrometer are also provided. The experiments are demonstrated for the protein calmodulin (16.7kDa). |
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Authors:
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Lewis E Kay; Mitsuhiko Ikura; Rolf Tschudin; Ad Bax |
Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Journal of magnetic resonance (San Diego, Calif. : 1997) Volume: 213 ISSN: 1096-0856 ISO Abbreviation: J. Magn. Reson. Publication Date: 2011 Dec |
Date Detail:
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Created Date: 2011-12-14 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9707935 Medline TA: J Magn Reson Country: United States |
Other Details:
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Languages: eng Pagination: 423-41 Citation Subset: IM |
Copyright Information:
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Copyright © 2011. Published by Elsevier Inc. |
Affiliation:
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Laboratory of Chemical Physics, NIDDK, National Institutes of Health, Bethesda, MD 20892, United States. |
Export Citation:
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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