TROSY NMR with partially deuterated proteins

TROSY-type optimization of liquid-state NMR experiments is based on the preservation of unique coherence transfer pathways with distinct transverse relaxation properties. The broadband decoupling of the ¹H spins interchanges the TROSY and anti-TROSY magnetization transfer pathways and thus is not used in TROSY-type triple resonance experiments or is replaced with narrowband selective decoupling. To achieve the full advantage of TROSY, the uniform deuteration of proteins is usually required. Here we propose a new and general method for ¹H broadband decoupling in TROSY NMR, which does not compromise the relaxation optimization in the ¹⁵N–¹H moieties, but uniformly and efficiently refocuses the ¹J_CH scalar coupling evolution in the ¹³C–¹H moieties. Combined with the conventional ²H decoupling, this method enables obtaining high sensitivity TROSY-type triple resonance spectra with partially deuterated or fully protonated ¹³C,¹⁵N labeled proteins.