Parallel acquisition of 3D-HA(CA)NH and 3D-HACACO spectra

We present here an NMR pulse sequence with 5 independent incrementable time delays within the frame of a 3-dimensional experiment, by incorporating polarization sharing and dual receiver concepts. This has been applied to directly record 3D-HA(CA)NH and 3D-HACACO spectra of proteins simultaneously using parallel detection of (1)H and (13)C nuclei. While both the experiments display intra-residue backbone correlations, the 3D-HA(CA)NH provides also sequential 'i - 1 → i' correlation along the (1)Hα dimension. Both the spectra contain special peak patterns at glycine locations which serve as check points during the sequential assignment process. The 3D-HACACO spectrum contains, in addition, information on prolines and side chains of residues having H-C-CO network (i.e., (1)Hβ, (13)Cβ and (13)COγ of Asp and Asn, and (1)Hγ, (13)Cγ and (13)COδ of Glu and Gln), which are generally absent in most conventional proton detected experiments.