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 1JCH 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.
Eletsky
A
A
Laboratorium für Physikalische Chemie, Eidgenossische Technische Hochschule Hönggerberg, Zürich, Switzerland.
Kienhöfer
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A
Pervushin
K
K
D016880:Anisotropy; D001412:Bacillus subtilis / Q000201:enzymology; D002247:Carbon Isotopes; D002826:Chorismate Mutase / Q000737:chemistry; D003903:Deuterium; D006859:Hydrogen; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins / Q000737:chemistry*
D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D011506:Proteins; D006859:Hydrogen; D003903:Deuterium; D002826:Chorismate Mutase