Untersuchungen zur Rolle der Tripeptidylpeptidase II-vermittelten Proteolyse invivo (In vivo-Role of Tripeptidyl Peptidase II-Mediated Proteolysis in Murine Cells and Mice)

三肽基肽酶 II 介导的蛋白水解作用的体内研究（In vivo-Role of Tripeptidyl Peptidase II-Mediated Proteolysis in Murine Cells and Mice）

• 批准号: 5111671
• 负责人: Professorin Dr. Gabriele Niedermann
• 金额: --
• 依托单位: Klinik für Strahlenheilkunde
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 1998
• 资助国家: 德国
• 起止时间: 1997-12-31 至 2005-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5111671?language=en
• 关键词: Untersuchungen; zur; Rolle; der; Tripeptidylpeptidase

Whereas some groups favor that proteasomes only generate the C-termini of cytotoxic T-lymphocyte (CTL) epitopes (Craiu et al., 1997; Beninga et al., 1998), our work has suggested that they cleave precisely at both ends of many epitopes. We intend to use as substrate HIV-1-Nef, which contains at least 45 CTL epitopes, for digestion by isolated 20S, 26S and immunoproteasomes and to determine all degradation products. This will allow us to clarify the role of proteasomes in epitope generation, to better define the specificity of the various types of proteasomes (the specificities of 26S and immunoproteasomes have so far not been investigated systematically) and to develop algorithms for the prediction of proteasomal cleavage sites.Upregulation of TPPII, a cytosolic protease complex larger than the 26S proteasome, in proteasome inhibitor-adapted cells permits cell survival. We intend to investigate whether TPPII normally functions in cytosolic proteolysis down-stream of proteasomes and therefore is the eukaryotic pendant of the archaeal Tricorn protease. Further, we intend to analyze the adapted cells in more detail regarding impairment of proteasome functions and precise mechanisms by which TPPII permits survival.

尽管有些群体倾向于蛋白酶体只产生细胞毒性T淋巴细胞(CTL)表位的C末端(Craiu等人，1997；Beninga等人，1998)，但我们的工作表明，它们恰好在许多表位的两端切割。我们打算使用HIV-1-Nef作为底物，它至少包含45个CTL表位，用于被分离的20S、26S和免疫蛋白酶体消化，并测定所有降解产物。这将使我们能够澄清蛋白酶体在表位生成中的作用，更好地定义各种类型的蛋白酶体的特异性(到目前为止，26S和免疫蛋白酶体的特异性还没有得到系统的研究)，并开发出预测蛋白酶体裂解位点的算法。在蛋白酶体抑制剂适应的细胞中，TPPII是一种比26S蛋白酶体大的胞浆蛋白酶复合体，上调TPPII使细胞存活。我们打算研究TPPII是否正常地在蛋白酶体下游的胞浆蛋白分解中发挥作用，从而是古代三角兽蛋白酶的真核悬挂。此外，我们打算更详细地分析适应的细胞蛋白酶体功能的损伤和TPPII允许生存的确切机制。