Nuclear magnetic resonance studies of flexible opiate conformations at monoclonal antibody binding sites. Quantitative interproton distances obtained from comparing theoretical and experimental transferred nuclear Overhauser enhancement: correlation with
Nuclear magnetic resonance studies of flexible opiate conformations at monoclonal antibody binding sites. Quantitative interproton distances obtained from comparing theoretical and experimental transferred nuclear Overhauser enhancement: correlation with
复制标题
单克隆抗体结合位点柔性阿片构象的核磁共振研究。
DOI：
10.1016/0022-2836(89)90604-9
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发表时间：
1989
影响因子：
5.6
通讯作者：
Glasel,JA
中科院分区：
文献类型：
--
作者：
Glasel,JA
以前的出版物描述了使用定性的intramolecular 1H-转移核Overhauser效应测量，以确定在单克隆抗阿片抗体结合位点的柔性配体的构象。本文集中在定量解释这种类型的实验，使用配体纳洛啡（N-烯丙基吗啡）和一个单一的抗阿片类药物的单克隆抗体。我比较实验一维驱动核Overhauser效应积累曲线与理论曲线得出的知识的固定的配体中的质子间距离。讨论涵盖了潜在的准确性派生的距离，并集中在两个问题领域与确定结构，从这种类型的实验。最严重的一种情况是，由于特定的多质子空间分布，自旋扩散是如此之快，以至于无法通过实验确定，并且理论计算的数值拟合是误导性的。结果表明，虽然配体内自旋扩散使某些质子对的解释复杂化，但对于2到4 μ m的质子间距离，可以达到约0.3 μ m的精度。结果证实了早先的报告，即在该抗体结合位点的纳洛啡的构象不同于溶液或晶体中存在的主要构象。一个重要的方面的工作是，理论预测的核Overhauser效应的时间依赖性是一个重要的实用工具，用于识别的情况下，解释的实验将是困难的。初始数据蛋白质到配体转移核Overhauser效应，这表明，至少有一个芳香族氨基酸残基密切参与的配体的结合。配套文件介绍了在我们的研究中使用的抗体的可变区的主要序列。在本文中，这些和相关的免疫化学研究与核磁共振结果。在这两篇论文中提出的数据的组合提供了一个基础，为未来的工作蛋白质配体质子间距离在1至5毫米的范围内使用转移核奥弗豪泽效应（快速交换配体）和同位素编辑，间接检测核奥弗豪泽效应（紧密结合的配体）。
A previous publication described the use of qualitative intramolecular1H-transferred nuclear Overhauser effect measurements to determine the conformations of flexible ligands at monoclonal anti-opiate antibody binding sites. This paper concentrates on the quantitative interpretation of experiments of this type using the ligand nalorphine (N-allyl morphine) and a single anti-opiate monoclonal antibody. I compare the experimental unidimensional driven nuclear Overhauser effect buildup curves to theoretical curves derived with a knowledge of the fixed interproton distances in the ligand. The discussion covers the potential accuracies of derived distances and concentrates on two problem areas associated with determining structures from this type of experiment. The most serious one is the case where, because of particular multiproton spatial distributions, spin diffusion is so rapid that it cannot be determined experimentally and where numerical fits of theoretical calculations are misleading. The results show that, while intraligand spin diffusion complicates the interpretation for some proton pairs, with many others accuracies within about 0·3 Å for interproton distances from 2 to 4 Å are attainable. The results confirm the earlier report that the conformation of nalorphine in this antibody binding site differs from the major one present in solution or in the crystal. An important aspect of the work is that theoretical prediction of nuclear Overhauser effect time-dependence is an important practical tool for recognizing cases where interpretation of experiments will be difficult.Initial data on protein-to-ligand transferred nuclear Overhauser effect are presented, which show that at least one aromatic amino acid residue is closely involved in the binding of the ligand. The companion paper presents the primary sequences of the variable regions of the antibodies being used in our studies. In this paper, these and associated immunochemical studies are correlated with the nuclear magnetic resonance results. The combination of data presented in the two papers provides a basis for future work on protein-ligand interproton distances in the range 1 to 5 Å using both transferred nuclear Overhauser effect (for rapidly exchanging ligands) and isotope-edited, indirectly detected nuclear Overhauser effect (for tightly bound ligands).