Structural and biochemical studies elucidating the biogenesis pathway of 20S proteasomes

结构和生化研究阐明 20S 蛋白酶体的生物发生途径

• 批准号: 442219341
• 负责人: Professor Dr. Jürgen Dohmen
• 金额: --
• 依托单位: Institut für Genetik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/442219341?language=en
• 关键词: Structural; biochemical; studies; elucidating; biogenesis

In this proposal we address the spatial and temporal order of events in the chaperone-guided assembly and maturation of proteasomal 20S core particle (CP) complexes. We aim to structurally characterise the late steps of CP biogenesis by determining high-resolution cryo EM reconstructions in order to identify the exact interactions between proteasomal chaperones and 20S CP subunits. At the same time, we want to analyse by which mechanism(s) autocatalytic activation of the beta subunits is initiated. In parallel, we want to investigate the so far poorly understood early steps of proteasome CP biogenesis biochemically and structurally through the characterization of individual assembly intermediates, which will moreover be used to recapitulate distinct assembly steps in vitro with the aim to determine the order of events and their checkpoints.

在这个建议中，我们解决的空间和时间顺序的事件在伴侣蛋白引导的组装和成熟的蛋白酶体20 S核心颗粒（CP）复合物。我们的目标是通过确定高分辨率cryo EM重建，以确定蛋白酶体伴侣和20 S CP亚基之间的确切相互作用，从结构上确定CP生物发生的后期步骤。与此同时，我们希望分析β亚基的自催化激活是通过何种机制启动的。与此同时，我们要调查迄今为止知之甚少的蛋白酶体CP生物合成的早期步骤的生化和结构通过表征的个人组装中间体，这将被用来概括不同的组装步骤在体外的目的是确定事件的顺序和他们的检查点。