CONFORMATIONS OF LIGANDS AT ANTIBODY BINDING SITES

这项研究将介绍当代单抗提供的工具 抗体(Mab)技术在配基-大分子物理研究中的应用 有约束力的。结构上大量使用的有利条件 以Fab片段的形式提供的均一抗体结合部位 克隆免疫球蛋白构成了这项工作的基础。我们计划 旨在阐明亚分子的一系列实验 一些具有生物学功能的半抗原与单抗的结合 针对它们和它们的结构类似物产生的抗体。两个人 初步研究选择的触觉系统是阿片类药物和 趋化性三肽。在前期工作中，我们获得了4条线路 针对重要药理表位的单抗 在吗啡上，并证明了它们的低解离常数。 吗啡及其与阿片激动剂和阿片类药物的不同交叉反应 对抗者。我们的一位合作者在 趋化三肽系统和这个项目现在已经被接管到 我们的实验室。所选择的物理方法是高频核 溶液研究的磁共振和中子衍射法 结晶的半抗原-Fab复合体。在解决方案中，使用来自 微观酸度常数、化学位移和质子-质子核 半抗原和免疫球蛋白共振的Overhauser效应 我们将形成连接免疫球蛋白残基的邻近图 与半抗原上的原子一起折叠。借助于特制的氢化物 半抗原的衍生物我们将确定它们的构象 用差中子衍射法研究自身结合位和交叉反应结合位 在晶体复合体上进行的。这项工作将与 LIGH和LIGH高变区的氨基酸序列分析 免疫球蛋白片段的重链。标记、衍射法和 每项排序工作都将与Expert合作完成 调查人员。 这项工作将与以前关于大分子的工作有很大的不同 结合部位。我们将致力于生物功能配体的研究 其中Mab工具允许我们检查 以详细的方式介绍大分子结合部位。此外，还有一个好的方面 在几个系统中出现的证据，包括趋化作用 多肽一，抗体结合部位可能是很好的模型 细胞受体结合部位呈现给半抗原，例如我们将被 在它们正常的生物功能过程中进行研究。

This research will introduce tools offered by contemporary monoclonal antibody (Mab) technology into physical studies of ligand-macromolecular binding. The advantageous use of the large quantities of structurally homogeneous antibody binding sites available in the form of Fab fragments of monoclonal immunoglobulins forms the basis of the work. We plan a series of experiments which are aimed at elucidating, in submolecular detail, the binding of some biologically functional haptens to monoclonal antibodies raised against them and their structural analogs. The two haptenic systems chosen for initial study are the opiates and the chemotactic tripeptides. In preliminary work we have obtained 4 lines of monoclonal antibodies directed against pharmacologically important epitopes on morphine and have demonstrated their low dissociation constants from morphine and their differential cross-reactivities to opiate agonists and antagonists. One of our collaborators has done similar work in the chemotactic tripeptide system and this project has now been taken over into our laboratory. The physical methods of choice are high frequency nuclear magnetic resonance for solution studies and neutron diffraction for crystallized hapten-Fab complexes. In solution, using information from microscopic acidity constants, chemical shifts and proton-proton Nuclear Overhauser Effects for resonances from both haptens and immunoglobulin fragments we will form proximity maps connecting residues in immunoglobulin folds with atoms on the haptens. With the aid of specifically deuterated derivatives of the haptens we will determine their conformations at their own and cross-reacting binding sites using difference neutron diffraction performed on crystalline complexes. This work will be correlated with amino acid sequence analysis of the hypervariable regions of the light and heavy chains of the immunoglobulin fragments. Labeling, diffraction and sequencing work will each be done in collaboration with expert investigators. This work will differ significantly from previous work on macromolecular binding sites. We will be working with biologically functional ligands where the Mab tool allows us to examine the fine structures of the macromolecular binding sites in a detailed way. In addition, there is good evidence which is emerging in several systems, including the chemotactic peptide one, that antibody binding sites may be good models for the cellular receptor binding sites presented to haptens, such as we will be studying during their normal biological functions.