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

我们在这个项目中的目标一直是研究 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/多肽复合体结构的计算机模拟。 这样的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，这是 在启动免疫反应中起着核心作用。在……里面 此外，它们为理解更大的问题提供了一个框架 蛋白质是如何与小分子多肽配体结合的。 过去一年的主要成就包括进一步发展 以及定量动力学分析的应用，用于测量 多肽与纯化的MHC分子的实时相互作用 对大量多肽结合的广泛分析。在……里面 此外，还设计了定量排空MHC的方法 I类分子及其多肽结合活性的保留。 此外，正在开发化学物质的一般方法。 标记有组氨酸残基的工程分子与金属的偶联- 共轭生物传感器表面。

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.