当前位置: X-MOL 学术J. Biomol. NMR › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Two-dimensional NMR lineshape analysis of single, multiple, zero and double quantum correlation experiments.
Journal of Biomolecular NMR ( IF 2.7 ) Pub Date : 2020-01-08 , DOI: 10.1007/s10858-019-00297-7
Christopher A Waudby 1 , Margaux Ouvry 2 , Ben Davis 3 , John Christodoulou 1
Affiliation  

NMR spectroscopy provides a powerful approach for the characterisation of chemical exchange and molecular interactions by analysis of series of experiments acquired over the course of a titration measurement. The appearance of NMR resonances undergoing chemical exchange depends on the frequency difference relative to the rate of exchange, and in the case of one-dimensional experiments chemical exchange regimes are well established and well known. However, two-dimensional experiments present additional complexity, as at least one additional frequency difference must be considered. Here we provide a systematic classification of chemical exchange regimes in two-dimensional NMR spectra. We highlight important differences between exchange in HSQC and HMQC experiments, that on a practical level result in more severe exchange broadening in HMQC spectra, but show that complementary alternatives to the HMQC are available in the form of HZQC and HDQC experiments. We present the longitudinal relaxation optimised SOFAST-H(Z/D)QC experiment for the simultaneous acquisition of sensitivity-enhanced HZQC and HDQC spectra, and the longitudinal and transverse relaxation optimised BEST-ZQ-TROSY for analysis of large molecular weight systems. We describe the application of these experiments to the characterisation of the interaction between the Hsp90 N-terminal domain and a small molecule ligand, and show that the independent analysis of HSQC, HMQC, HZQC and HDQC experiments provides improved confidence in the fitted dissociation constant and dissociation rate. Joint analysis of such data may provide improved sensitivity to detect and analyse more complex multi-state interaction mechanisms such as induced fit or conformational selection.

中文翻译:

单量子、多量子、零量子和双量子相关实验的二维核磁共振线形分析。

核磁共振波谱通过分析滴定测量过程中获得的一系列实验,为表征化学交换和分子相互作用提供了一种强大的方法。经历化学交换的 NMR 共振的出现取决于相对于交换速率的频率差,并且在一维实验的情况下,化学交换机制是众所周知的。然而,二维实验呈现出额外的复杂性,因为必须考虑至少一个额外的频率差。在这里,我们提供了二维核磁共振谱中化学交换机制的系统分类。我们强调了 HSQC 和 HMQC 实验中交换之间的重要差异,这在实际水平上导致 HMQC 光谱中更严重的交换展宽,但表明可以以 HZQC 和 HDQC 实验的形式提供 HMQC 的补充替代方案。我们提出了纵向弛豫优化的 SOFAST-H(Z/D)QC 实验,用于同时采集灵敏度增强的 HZQC 和 HDQC 光谱,以及纵向和横向弛豫优化的 BEST-ZQ-TROSY,用于分析大分子量系统。我们描述了这些实验在表征 Hsp90 N 端结构域和小分子配体之间相互作用的应用,并表明 HSQC、HMQC、HZQC 和 HDQC 实验的独立分析提高了拟合解离常数和解离常数的可信度。解离率。对此类数据的联合分析可以提高检测和分析更复杂的多状态相互作用机制(例如诱导拟合或构象选择)的灵敏度。
更新日期:2020-01-08
down
wechat
bug