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 H-1 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 H-1 broadband decoupling in TROSY NMR, which does not compromise the relaxation optimization in the N-15-H-1 moieties, but uniformly and efficiently refocuses the (1)J(CH) scalar coupling evolution in the C-13-H-1 moieties. Combined with the conventional H-2 decoupling, this method enables obtaining high sensitivity TROSY-type triple resonance spectra with partially deuterated or fully protonated C-13,N-15 labeled proteins.