Implications of ß-subunit exchanges and of interactions with the activator PA28 for the cleavage preferences of the 20S proteasome: A quantitative analysis based on kinetic experiments

× 亚基交换以及与激活剂 PA28 的相互作用对 20S 蛋白酶体切割偏好的影响：基于动力学实验的定量分析

• 批准号: 5252194
• 负责人: Professor Dr. Hermann-Georg Holzhütter
• 金额: --
• 依托单位: Institut für Biochemie (CCM)
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2000
• 资助国家: 德国
• 起止时间: 1999-12-31 至 2005-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5252194?language=en
• 关键词: Implications; subunit; exchanges; interactions; activator

The structural and kinetic features accounting for the highly selective processing of protein substrates by eukaryotic 20S proteasomes are still poorly understood. The research proposal aims at a comprehensive kinetic modelling of the 20S proteasome. This enables one to test by computer simulations how various putative cleavage mechanisms will affect the pattern of cleavage fragments derived from a given protein substrate. Comparing the model predictions with experimentally determined patterns of proteol ytic fragments the most probable cleavage mechanism will eventually be figured out. To this end an appropriate data base will be established digesting various oligomeric substrates under strictly controlled external conditions (proteasome preparation, pre-treatment of substrates, presence of activating agents, time points etc.) by the constitutive and 'immuno' 20S proteasome and recording the time-dependent formation of the major cleavage products. It is planned to apply the kinetic model as a predictor of major proteasomal cleavage products to scan arbitrary protein sequences for the presence of potential antigenic epitopes.

真核20 S蛋白酶体对蛋白质底物的高选择性加工的结构和动力学特征仍然知之甚少。该研究计划旨在对20S蛋白酶体进行全面的动力学建模。这使得人们能够通过计算机模拟来测试各种假定的切割机制将如何影响来自给定蛋白质底物的切割片段的模式。将模型预测与实验确定的蛋白水解片段模式进行比较，最终将找出最可能的切割机制。为此，将建立适当的数据库，在严格控制的外部条件（蛋白酶体制备、底物预处理、活化剂的存在、时间点等）下消化各种寡聚底物。通过组成型和“免疫”20 S蛋白酶体，并记录主要裂解产物的时间依赖性形成。计划应用动力学模型作为主要蛋白酶体切割产物的预测因子，以扫描任意蛋白质序列中潜在抗原表位的存在。