Sequential protein NMR assignments in the liquid state via sequential data acquisition

Two different NMR pulse schemes involving sequential 1 H data acquisition are presented for achieving protein backbone sequential resonance assignments: (i) acquisition of 3D {HCCNH and HNCACONH} and (ii) collection of 3D {HNCOCANH and HNCACONH} chemical shift correlation spectra using uniformly 13 C, 15 N labelled proteins. The sequential acquisition of these spectra reduces the overall experimental time by a factor of ≈ 2 as compared to individual acquisitions. The suitability of this approach is experimentally demonstrated for the C-terminal winged helix (WH) domain of the minichromosome maintenance (MCM) complex of Sulfolobus solfataricus .