IN VITRO FOLDING AND ASSEMBLY OF MHC MOLECULES

这个项目的目的一直是了解分子机制。 控制MHC I类分子的组装和折叠。这个 MHC I类分子作为关键的细胞表面结构 用于识别的细胞内肽片段样本 抗原特异的MHC限制性T淋巴细胞。在这个项目中，我们有 试图在体外系统中理解这种成分是如何 由MHC I类分子、多肽、 而轻链，β2-微球蛋白遵循的途径是从它们的 作为单个细胞表面分子的分离链的生物合成。通过 设计这些分子的单链类似物以表达两者 在细菌和组织培养细胞中，我们已经能够检查 多肽和轻链成分对最终产物的贡献 结构。这使得我们能够为生产制定一个战略 只表达一种I类MHC分子并产生 表达单链I类分子的转基因动物。这些 研究不仅应该让我们了解蛋白质的基本规则 折叠，因为它们属于三聚体MHC I类分子，但也 允许我们利用这一知识来生成动物模型 高度受限的MHC-I类表达。

The aim of this project has been to understand the molecular mechanisms that control the assembly and folding of the MHC class I molecules. The MHC Class I molecules serve as the critical cell surface structures that sample intracellular peptide fragments for recognition by antigen-specific, MHC-restricted T lymphocytes. In this project we have sought to understand in an in vitro system how the components of this three chain structure consisting of the MHC class I molecule, peptide, and the light chain, beta2-microglobulin follow a pathway from their biosynthesis as separate chains to a single cell surface molecule. By engineering single chain analogues of these molecules for expression both in bacteria and in tissue culture cells, we have been able to examine the contribution of the peptide and light chain components to the final structure. This has allowed us to develop a strategy for the production of cell lines that express only one class I MHC molecule and to generate transgenic animals that express single chain class I molecules. These studies should allow us not only to understanding basic rules of protein folding as they pertain to the trimer MHC class I molecule, but also allow us to exploit this knowledge in generating animal models with highly restricted MHC class I expression.