TROSY-based HNCO pulse sequences for the measurement of (1)HN-(15)N, (15)N-(13)CO, (1)HN-(13)CO, (13)CO-(13)C(α) and (1)HN-(13)C(α) dipolar couplings in (15)N, (13)C, (2)H-labeled proteins

HNCO-based 3D pulse schemes are presented for measuring 1HN-15N,15N-13CO, 1HN-13CO,13CO-13C(α) and 1HN-13C(α) dipolar couplings in 15N,13C,2H-labeled proteins. The experiments are based on recently developed TROSY methodology for improving spectral resolution and sensitivity. Data sets recorded on a complex of Val, Leu, Ile (δ1 only) methyl protonated 15N,13C,2H-labeled maltose binding protein and β-cyclodextrin as well as 15N,13C,2H-labeled human carbonic anhydrase II demonstrate that precise dipolar couplings can be obtained on proteins in the 30-40 kDa molecular weight range. These couplings will serve as powerful restraints for obtaining global folds of highly deuterated proteins.

Duke Scholars

Author Ronald Venters Radiology