Cell surface display of functional human MHC class II proteins: yeast display versus insect cell display.
Cell surface display of functional human MHC class II proteins: yeast display versus insect cell display.
复制标题
DOI：
10.1093/protein/gzr035
复制
发表时间：
2011-09
期刊：
影响因子：
--
通讯作者：
Fei Wen;D. Sethi;K. Wucherpfennig;Huimin Zhao
中科院分区：
文献类型：
--
作者：
Fei Wen;D. Sethi;K. Wucherpfennig;Huimin Zhao
工程功能性主要组织相容性复合体II类(MHCII)蛋白的可靠和健壮的系统已被证明是难以捉摸的。这些系统的可用将使多肽-MHCII(PMHCII)复合体的工程应用于治疗和诊断应用。在这篇论文中，我们开发了一种基于昆虫细胞表面展示的系统，该系统允许带有或不带有共价结合的人髓鞘碱性蛋白(MBP)肽的异源二聚体DR2分子的功能性表达，这符合DR2-MBP变体的定向进化，具有更好的T细胞受体(TCR)结合亲和力。本研究首次在昆虫细胞表面展示了人pMHCII复合体的功能。在开发这一pMHCII工程系统的过程中，我们还探索了将酵母表面展示用于相同应用的潜力。我们的数据表明，酵母展示是一个有用的系统来分析和设计MHCII蛋白的肽结合，但不适合于与自身反应性TCR结合的低亲和力的pMHC复合体的定向进化。
Reliable and robust systems for engineering functional major histocompatibility complex class II (MHCII) proteins have proved elusive. Availability of such systems would enable the engineering of peptide-MHCII (pMHCII) complexes for therapeutic and diagnostic applications. In this paper, we have developed a system based on insect cell surface display that allows functional expression of heterodimeric DR2 molecules with or without a covalently bound human myelin basic protein (MBP) peptide, which is amenable to directed evolution of DR2-MBP variants with improved T cell receptor (TCR)-binding affinity. This study represents the first example of functional display of human pMHCII complexes on insect cell surface. In the process of developing this pMHCII engineering system, we have also explored the potential of using yeast surface display for the same application. Our data suggest that yeast display is a useful system for analysis and engineering of peptide binding of MHCII proteins, but not suitable for directed evolution of pMHC complexes that bind with low affinity to self-reactive TCRs.