DEPT spectral editing in HCCONH-type experiments. Application to fast protein backbone and side chain assignment.
2D DEPT-H(alpha,beta)C(alpha,beta)(CO)NH and 2D CT-DEPT-HC(CO)NH-TOCSY experiments are presented which allow fast resonance assignment of aliphatic protein side chains. In these 2D reduced-dimensionality experiments, two or three nuclei are frequency labeled in the indirect dimension. DEPT spectral editing reduces the number of correlation peaks detected in each 2D spectrum, and helps in amino-acid-type determination during sequential backbone resonance assignment. Applications are shown for a small 68-residue, and a highly deuterated 167-residue protein. The new experiments complement the set of 2D HNX correlation experiments, previously proposed for fast protein resonance assignment [J. Biomol. NMR, 27 (2003) 57].
Brutscher
Bernhard
B
Institut de Biologie Structurale-Jean-Pierre Ebel, UMR5075 CNRS-CEA-UJF, 41, rue Jules Horowitz-38027, Grenoble Cedex 1, France. Bernhard.Brutscher@ibs.fr
D000465:Algorithms; D000595:Amino Acid Sequence; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000032:analysis* / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000032:analysis / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012815:Signal Processing, Computer-Assisted
D011506:Proteins; D010088:Oxidoreductases; C021551:mercuric reductase