Saturation-Transfer Difference (STD) NMR: A Simple and Fast Method for Ligand Screening and Characterization of Protein Binding
Saturation-Transfer Difference (STD) NMR: A Simple and Fast Method for Ligand Screening and Characterization of Protein Binding
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DOI：
10.1021/ed101169t
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发表时间：
2011-07-01
影响因子：
3
通讯作者：
Cabrita, Eurico J.
中科院分区：
文献类型：
--
作者：
Viegas, Aldino;Manso, Joao;Cabrita, Eurico J.
饱和转移差(STD)核磁共振已成为研究蛋白质配基相互作用的最常用的基于配基的核磁共振技术之一。这种技术的成功是因为它的健壮性和它专注于配体的信号，而不需要处理关于受体的核磁共振信息，只使用少量的非标记大分子。此外，这个实验的吸引力也可以延伸到课堂上。在一个实用的核磁共振课程的背景下，这个实验是理想的说明一些基本的核磁共振概念，如多学科背景下的核Overhauser效应和弛豫，将化学和生物化学与药物化学的味道联系起来。我们使用容易获得的人血清白蛋白，6-D，L-甲基色氨酸(6-CH(3)-Trp)和7-D，L-甲基色氨酸(7-CH(3)-Trp)来介绍STD-核磁共振实验，并说明它在配体筛选，结合部分的映射和离解常数测定的适用性，在一个可以探索或适应学生的课程水平和主题(化学或生物化学)的背景下。我们还讨论了STD实验的最重要的理论方面，提醒人们注意它的一些限制和缺陷，而不是复杂的理论方法。
Saturation transfer difference (STD) NMR has emerged as one of the most popular ligand-based NMR techniques for the study of protein ligand interactions. The success of this technique is a consequence of its robustness and the fact that it is focused on the signals of the ligand, without any need of processing NMR information about the receptor and only using small quantities of nonlabeled macromolecule. Moreover, the attractiveness of this experiment is also extendable to the classroom. In the context of a practical NMR class, this experiment is ideal to illustrate some fundamental NMR concepts, such as the nuclear Overhauser effect and relaxation in a multidisciplinary context, bridging chemistry and biochemistry with a taste of medicinal chemistry. We use the readily available human serum albumin (HSA), 6-D,L-methyl-tryptophan (6-CH(3)-Trp), and 7- D,L-methyl-tryptophan (7-CH(3)-Trp) to introduce the STD-NMR experiment and to illustrate its applicability for ligand screening, mapping of binding moieties, and determination of the dissociation constant, in a context that can be explored or adapted to the student's course level and topic (chemistry or biochemistry). We also cover the most important theoretical aspects of the STD experiment, calling attention to some of its limitations and drawbacks without a complex theoretical approach.