A simple and sensitive experiment for measurement of JCC couplings between backbone carbonyl and methyl carbons in isotopically enriched proteins |
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Authors: | Stephan Grzesiek Geerten W Vuister Ad Bax |
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Institution: | (1) Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, 20892 Bethesda, MD, U.S.A. |
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Abstract: | Summary A simple 2D difference experiment is described that allows quantitative measurement of 13C–13C J couplings between backbone carbonyl and side-chain carbons. Precise 3JCC values were measured from data recorded in just 2 h for a 1-mM solution of the 20-kD complex between the protein calmodulin and a 26-residue synthetic peptide. The J couplings aid in determining the 1 angles of valine, isoleucine and threonine residues, and in making stereospecific assignments of the Val C methyl groups. Error analysis indicates that the uncertainty in the derived J couplings is generally less than ca. 0.3 Hz. |
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Keywords: | Carbon-carbon J coupling Proteins Two-dimensional NMR 1 Angle" target="_blank">gif" alt="chi" align="MIDDLE" BORDER="0">1 Angle Stereospecific assignment |
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