Studying MHC class I peptide loading and exchange in vitro.
Studying MHC class I peptide loading and exchange in vitro.
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研究 MHC I 类肽的体外装载和交换。
DOI：
10.1007/978-1-62703-218-6_7
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Bouvier,Marlene
中科院分区：
文献类型：
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作者：
Bouvier,Marlene
在内质网（ER）中，MHC I类分子与几种专门的蛋白质结合，形成称为“肽加载复合物”（PLC）的大分子复合物。在PLC中，抗原肽经历严格的选择过程以结合到MHC I类分子上。这确保了免疫系统对病毒和实体瘤产生强大的CD8+ T细胞应答。在体外重构与PLC的关键蛋白相关的MHC I类分子的能力提供了在分子水平上研究如何选择抗原肽以呈递给CD8+ T细胞的手段。在这里，我们描述了用于产生涉及MHC I类分子和塔帕辛，PLC的一个关键蛋白质的无细胞系统的实用程序，它可以用作肽装载和交换的生化和机械研究的通用工具。
In the endoplasmic reticulum (ER), MHC class I molecules associate with several specialized proteins, forming a large macromolecular complex referred to as the “peptide-loading complex” (PLC). In the PLC, antigenic peptides undergo a stringent selection process for binding onto MHC class I molecules. This ensures that the immune system elicits robust CD8+ T-cell responses to viruses and solid tumors. The ability to reconstitute in vitro MHC class I molecules in association with key proteins of the PLC provides a mean for studying at the molecular level how antigenic peptides are selected for presentation to CD8+ T-cells. Here, we describe practical procedures for generating a cell-free system involving MHC class I molecules and tapasin, a critical protein of the PLC, that can be used as a versatile tool for biochemical and mechanistic studies of peptide loading and exchange.