Semi-constant-time HMSQC (SCT-HMSQC-HA) for the measurement of (3)J(H(N))(H(alpha)) couplings in (15)N-labeled proteins.
A simple method for accurately measuring (3)J(H(N))(H(alpha)) coupling constants in (15)N-labeled proteins is described. This semi-constant-time HMSQC-HA experiment combines the rapidity and convenience of the recently introduced CT-HMQC-HA scheme (Postingl and Otting, J. Biomol. NMR 12, 319-324 (1998)) with the high resolution and robustness of the HSQC experiment. The proposed method is demonstrated for the 76-residue human ubiquitin and Saccharopolyspora erythraea calerythrin (176 residues). Our results imply that the SCT-HMSQC-HA experiment is suitable also for proteins with less favorable NMR properties due to its good resolution and sensitivity.
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NMR Laboratory, Institute of Biotechnology, University of Helsinki, FIN-00014, Finland.
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D000596:Amino Acids / Q000032:analysis; D001426:Bacterial Proteins / Q000737:chemistry; D002135:Calcium-Binding Proteins / Q000737:chemistry; D006801:Humans; D006859:Hydrogen; D006860:Hydrogen Bonding; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D009587:Nitrogen Isotopes; D011506:Proteins / Q000737:chemistry*; D016930:Saccharopolyspora / Q000737:chemistry; D012680:Sensitivity and Specificity; D013113:Spin Labels; D013997:Time Factors; D014452:Ubiquitins / Q000737:chemistry
D000596:Amino Acids; D001426:Bacterial Proteins; D002135:Calcium-Binding Proteins; D009587:Nitrogen Isotopes; D011506:Proteins; D013113:Spin Labels; D014452:Ubiquitins; C090958:calerythrin protein, Saccharopolyspora erythraea; D006859:Hydrogen