NMR PROTON EXCHANGE STUDIES OF ANTIBODY CROSS-REACTIONS

抗体交叉反应的核磁共振质子交换研究

• 批准号: 3292457
• 负责人: PETER S KIM
• 金额: $ 11.14万
• 依托单位: WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-07-01 至 1989-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3292457
• 关键词: antibody; antibody formation; circular dichroism; conformation; deuterium; nonradiation isotope effect; nuclear magnetic resonance spectroscopy; peptides; radiotracer; trypsin inhibitors

The long-term objective of this proposed work is to establish a framework for using anti-peptide antibodies as probes of protein folding and dynamics. It is necessary first to investigate the mechanism of cross-reaction between intact proteins and anti-peptide antibodies. It is also necessary to develop a convenient method for studying the structures of proteins while they are bound to anti-peptide antibodies. We will study bovine pancreatic trypsin inhibitor (BPTI), a protein that has been well-characterized by biophysical and structural methods. Difference circular dichroism (CD) spectroscopy will be used to investigate the mechanism of cross-reaction at low resolution. The kinetics and thermodynamics of cross-reaction will be characterized. We will also determine if there is a correlation between the ability of peptides to fold, as judged by CD and nuclear magnetic resonance (NMR), and the ability of peptides to elicit cross-reacting antibodies. Amide proton exchange, combined with two-dimensional NMR spectroscopy (2D-NMR) will be used to study the structure of BPTI, while it is bound to anti-peptide antibodies. Hydrogen/deuterium exchange in the BPTI-antibody complex will be allowed to occur. Then, BPTI will be separated from antibody, and 2D-NMR will be used to quantitate the extent of exchange for individual amide protons in BPTI.

这项拟议工作的长期目标是建立一个框架 使用抗肽抗体作为蛋白质折叠的探针和 动力学。有必要首先研究其发病机制。 完整蛋白和抗肽抗体之间的交叉反应。它是 也有必要开发一种方便的方法来研究结构 当蛋白质与抗肽抗体结合时。我们会研究 牛胰腺胰蛋白酶抑制物(BPTI)，一种已经被 以生物物理和结构方法为特征的。 将使用差分圆二色谱(CD)来研究 低分辨率下的交叉反应机理。动力学和 交叉反应的热力学将被描述。我们还将 确定多肽的能力之间是否存在相关性 折叠，通过CD和核磁共振(核磁共振)判断，并能够 以引发交叉反应抗体。 酰胺质子交换，结合二维核磁共振波谱 (2D-核磁共振)将用于研究BPTI的结构，而它势必 抗肽抗体。BPTI抗体中的氢/氢交换 将允许发生复杂的情况。然后，BPTI将与 抗体，2D-核磁共振将被用来定量交换的程度 BPTI中的单个酰胺质子。