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).
Pang
Y
Y
Biophysics Research Division, University of Michigan, Ann Arbor 48109-1055, USA.
Zeng
L
L
Kurochkin
A V
AV
Zuiderweg
E R
ER
D001407:Bacillus / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D011487:Protein Conformation; D012260:Ribonucleases / Q000737:chemistry*
D012260:Ribonucleases