A suite of NMR pulse schemes is presented for the assignment of HN, 15N, 13Cα, and 13Cβ resonances in 15N, 13C, 2H labeled proteins. The experiments exploit the line narrowing of the 13C spins caused by the substitution of deuterons for bound protons and are therefore well suited for application to molecules with masses on the order of 30-40 kDa where standard triple resonance experiments may fail completely or provide spectra of poor sensitivity. The methods are demonstrated on a 37 kDa ternary complex of a 15N, 13C, and ~70% 2H labeled sample of tip-repressor, the corepressor 5-methyltryptophan, and a 20 basepair trp-operator DNA fragment. All of the experiments in the family presented provide well over 95% of the expected intra-and/or inter-residue correlations. Carbon relaxation measurements of the complex indicate that the use of deuteration increases the average 13Cα T2 value from 16.5 to 130 ms.
All Science Journal Classification (ASJC) codes
- Colloid and Surface Chemistry