IN VITRO FOLDING AND ASSEMBLY OF MHC MOLECULES

MHC 分子的体外折叠和组装

• 批准号: 3746610
• 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/3746610
• 关键词: MHC class I antigen; globulins; major histocompatibility complex; protein biosynthesis; protein engineering; protein folding; protein structure function; synthetic peptide

The aim of this project has been to develop in vitro systems for the study of the assembly and folding of the MHC class I molecules. Past studies have focused on coupled in vitro transcription/translation systems for examination of the assembly of the three chains of this macromolecule: the class I heavy chain, beta-2 microglobulin, and antigenic peptide. Recent results with in vitro mutants emphasize the value of such a system in evaluating parameters of assembly. Our current approach is to develop more biochemically defined methods, based on genetically engineered, bacterially produced single chain MHC class I proteins and synthetic antigenic peptides. We have demonstrated that biologically active, immunostimulatory MHC class I peptide complexes can be made by in vitro refolding of completely reduced and denatured molecules, suggesting that this is a feasible approach to produce large scale amounts of homogenous complexes for experimental use.

该项目的目的是开发体外系统， 研究MHC I类分子的组装和折叠。 过去 研究集中在体外转录/翻译偶联 系统的三个链的组装检查， 大分子：I类重链、β-2微球蛋白，以及 抗原肽 最近的体外突变体研究结果强调， 该系统在装配参数评价中的应用价值。 我们目前 方法是开发更多的生物化学定义的方法，基于 基因工程，细菌产生的单链MHC I类 蛋白质和合成抗原肽。 我们已经证明 具有生物活性的免疫刺激性MHC I类肽复合物可 通过体外重折叠完全还原和变性的 分子，这表明这是一种可行的方法，以产生大的 用于实验用途的均匀复合物的比例量。