Efficient measurement of (3)J(N,Cgamma) and (3)J(C',Cgamma) coupling constants of aromatic residues in (13)C, (15)N-labeled proteins.
An NMR pulse sequence is proposed for the simultaneous determination of side chain chi1 torsion-angle related (3)J(N,Cgamma) and (3)J(C', Cgamma) couplings in aromatic amino acid spin systems. The method is of the quantitative J correlation type and takes advantage of attenuated (15)N and (1)H transverse relaxation by means of the TROSY principle. Unlike previously developed schemes for the measurement of either of the two coupling types, spectra contain internal reference peaks that are usually recorded in separate experiments. Therefore, the desired information is extracted from a single rather than four data sets. The new method is demonstrated with uniformly (13)C/(15)N labeled Desulfovibrio vulgaris flavodoxin, which contains 14 aromatic out of 147 total amino acid residues.
Löhr
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Institut für Biophysikalische Chemie, Johann Wolfgang Goethe-Universität Frankfurt, Biozentrum N230, Marie Curie-Strasse 9, Frankfurt am Main, 60439, Germany.
Rüterjans
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D002247:Carbon Isotopes; D003901:Desulfovibrio / Q000737:chemistry; D005418:Flavodoxin / Q000737:chemistry; D009587:Nitrogen Isotopes; D010084:Oxidation-Reduction; D011506:Proteins / Q000737:chemistry*
D002247:Carbon Isotopes; D005418:Flavodoxin; D009587:Nitrogen Isotopes; D011506:Proteins