IN VITRO MEASUREMENT OF BINDING OF SELF AND ANTIGENIC PEPTIDES TO MHC MOLECULES

自身肽和抗原肽与 MHC 分子结合的体外测量

• 批准号: 3746609
• 负责人: D H MARGULIES
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3746609
• 关键词: MHC class I antigen; antigen antibody reaction; autoantigens; chemical binding; chemical kinetics; intermolecular interaction; major histocompatibility complex; mathematical model; protein engineering; protein structure function

Our objectives in this project have been to examine the details of the interaction of self and antigenic peptides with the MHC class I molecule by detailed kinetic and equilibrium binding methods, and by three- dimensional computer modeling of structures of MHC/peptide complexes. Such studies permit us to examine underlying biochemical rules that govern the interaction of antigenic peptides with MHC molecules, which plays a central role in the initiation of the immune response. In addition, they provide a framework for understanding the larger question of how proteins bind small molecular weight peptide ligands. Major accomplishments in the past year include the further development and application of quantitative kinetic assays for measurement of the interaction of peptides with purified MHC molecules in real time, and the extensive analysis of the binding of a larger number of peptides. In addition, methods have been devised for the quantitative emptying of MHC class I molecules and the retention of their peptide binding activity. In addition, general methods are being developed for the chemical coupling of engineered molecules tagged with histidine residues to metal- conjugated biosensor surfaces.

我们在这个项目中的目标是检查 自身和抗原肽与MHC I类分子的相互作用 通过详细的动力学和平衡结合方法，并通过三个- MHC/肽复合物结构的三维计算机建模。 这些研究使我们能够检查潜在的生化规则， 控制抗原肽与MHC分子的相互作用， 在免疫反应的启动中起着核心作用。 在 此外，它们为理解更大的问题提供了一个框架， 蛋白质如何结合小分子量的肽配体。 过去一年的主要成就包括： 和应用定量动力学分析来测量 肽与纯化的MHC分子在真实的时间内的相互作用， 大量肽结合的广泛分析。 在 此外，还设计了定量排空MHC的方法， I类分子和保留它们的肽结合活性。 此外，目前正在制定化学品的一般方法， 用组氨酸残基标记的工程化分子与金属- 共轭生物传感器表面。