High-resolution detection of five frequencies in a single 3D spectrum: HNHCACO--a bidirectional coherence transfer experiment.
A new triple-resonance pulse sequence, 3D HNHCACO, is introduced and discussed, which identifies sequential correlations of the backbone nuclei (H alpha (i-1), C alpha (i-1), C(i-1), NH(i), N(i)) of doubly labeled proteins in H2O. The three-dimensional (3D) method utilizes a recording of 15N and 13C resonances in a single indirect time domain, the 13C' resonance in another indirect time domain, and detects both NH and H alpha protons. A bidirectional coherence transfer (NH(i) <--> N(i) <--> C(i-1) <--> C alpha (i-1) <--> H alpha (i-1)) is effectuated, resulting in a single high-resolution 3D spectrum that contains the frequencies of all five backbone nuclei. The experiment was applied to the 12.3 kDa ribonuclease from Bacillus intermedius (Binase).
Biophysics Research Division, University of Michigan, Ann Arbor 48109-1055, USA.
Y
Y
Pang
L
L
Zeng
A V
AV
Kurochkin
E R
ER
Zuiderweg
D001407:Bacillus / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D011487:Protein Conformation; D012260:Ribonucleases / Q000737:chemistry*
D012260:Ribonucleases