Rosen Osnat, Anglister Jacob
Department of Structural Biology, Weizmann Institute of Science, Rehovot 76100, Israel.
Methods Mol Biol. 2009;524:37-57. doi: 10.1007/978-1-59745-450-6_3.
Nuclear magnetic resonance (NMR) is a very powerful tool for determining the boundaries of peptide epitopes recognized by antibodies. NMR can be used to study antibodies in complexes that exhibit a wide range of binding affinities from very weak and transient to very tight. Choice of the specific method depends upon the dissociation constant, especially the ligand off-rate.Epitope mapping by NMR is based on the difference in mobility between the amino acid residues of a peptide antigen that interact tightly with the antibody and residues outside the epitope that do not interact with the antibody. The interacting peptide residues become considerably immobilized upon binding. Their mobility will resemble that of the antibody's residues. Several NMR methods were developed based on these characteristics. In this chapter we discuss some of these methods, including dynamic filtering, comparison of (1)H-(15)N HSQC peaks' intensities, transverse relaxation time, measurements of (1)H-(15)N nuclear Overhauser effect (NOE) values, and measurements of T (1rho) relaxation time.
核磁共振（NMR）是确定抗体识别的肽表位边界的一种非常强大的工具。核磁共振可用于研究复合物中的抗体，这些复合物表现出从非常弱且短暂到非常紧密的广泛结合亲和力。具体方法的选择取决于解离常数，尤其是配体的解离速率。通过核磁共振进行表位作图是基于与抗体紧密相互作用的肽抗原氨基酸残基和表位外不与抗体相互作用的残基之间的迁移率差异。相互作用的肽残基在结合时会变得相当固定。它们的迁移率将类似于抗体的残基。基于这些特性开发了几种核磁共振方法。在本章中，我们将讨论其中一些方法，包括动态过滤、（1）H - （15）N HSQC峰强度比较、横向弛豫时间、（1）H - （15）N核Overhauser效应（NOE）值测量以及T（1rho）弛豫时间测量。
