Eletsky A, Kienhöfer A, Pervushin K
Laboratorium für Physikalische Chemie, Eidgenossische Technische Hochschule Hönggerberg, Zürich, Switzerland.
J Biomol NMR. 2001 Jun;20(2):177-80. doi: 10.1023/a:1011265430149.
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.
液态核磁共振实验的TROSY型优化基于具有独特横向弛豫特性的独特相干转移路径的保留。1H自旋的宽带去耦会互换TROSY和反TROSY磁化转移路径，因此在TROSY型三重共振实验中不使用，或者用窄带选择性去耦代替。为了充分利用TROSY的优势，通常需要对蛋白质进行均匀氘代。在此，我们提出了一种用于TROSY核磁共振中1H宽带去耦的新通用方法，该方法不会损害15N-1H部分的弛豫优化，但能均匀且有效地重新聚焦13C-1H部分中1JCH标量耦合的演化。结合传统的2H去耦，该方法能够获得具有部分氘代或完全质子化的13C、15N标记蛋白质的高灵敏度TROSY型三重共振谱。
