Analysis of the role of HSP100/Clp and their adaptor proteins in general and regulated proteolysis in Bacillus subtilis

分析 HSP100/Clp 及其衔接蛋白在枯草芽孢杆菌中的一般作用和调节蛋白水解作用

• 批准号: 5361999
• 负责人: Professor Dr. Kürsad Turgay
• 金额: --
• 依托单位: Max-Planck-Forschungsstelle für die Wissenschaft der Pathogene
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2002
• 资助国家: 德国
• 起止时间: 2001-12-31 至 2010-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5361999?language=en
• 关键词: Analysis; role; HSP100; Clp; their

Previously we could demonstrate that the adaptor protein MecA is necessary for the regulated proteolysis of the transcription factor ComK in competence development of Bacillus subtilis by ClpCP protease. However recent experimental data demonstrate that MecA is also necessary for the recognition, disaggregation and refolding of previously heat aggregated proteins by ClpC a chaperone of the HSP 100 subfamily of AAA+ ATPases. In addition MecA enables the specific recognition and degradation of unfolded or aggregated proteins by ClpCP. A MecA paralogue YpbH shows the same activity. The adaptor proteins are therefore necessary for the general function of ClpC in protein quality control in B. subtilis. To understand the role of the adaptor proteins MecA and the paralogue YpbH in the protein quality control system of B. subtilis we want to biochemically characterise and identify the partners of the adaptor proteins. Therefore we want to test the functional interaction of MecA and YpbH with those possible partner the HSP 100 proteins ClpC, ClpE and ClpX with and without ClpP. On the other hand we want to determine the general substrate specificity of the adaptor proteins, because the connection of substrate recognition and interaction with the HSP 100 is a prerequisite for the function of this new class of proteins. In addition experiments are devised to develop an understanding of the targeting for degradation mechanism of the adaptor proteins. Finally first experiments to understand the in vivo role of these adaptor proteins and their HSP 100 partner proteins in the protein quality control network of B. subtilis are introduced.

以前，我们可以证明，衔接蛋白MecA是必要的调节蛋白水解的转录因子ComK在感受态开发的枯草芽孢杆菌ClpCP蛋白酶。然而，最近的实验数据表明，MecA也是必要的识别，解聚和重折叠的先前热聚集的蛋白质由ClpC的AAA+ ATP酶的HSP 100亚家族的伴侣。此外，MecA能够通过ClpCP特异性识别和降解未折叠或聚集的蛋白质。MecA paralysis YpbH显示出相同的活性。因此，衔接蛋白对于B中ClpC在蛋白质质量控制中的一般功能是必需的。枯草芽孢杆菌了解接头蛋白MecA和接头蛋白YpbH在B蛋白质质量控制系统中的作用。枯草杆菌，我们想生化鉴定和确定配偶体的衔接蛋白。因此，我们想要测试MecA和YpbH与那些可能的伴侣HSP 100蛋白ClpC、ClpE和ClpX（有和没有ClpP）的功能性相互作用。另一方面，我们希望确定接头蛋白的一般底物特异性，因为底物识别和与HSP 100相互作用的连接是这类新蛋白功能的先决条件。此外，设计实验以发展对衔接蛋白降解机制的靶向理解。最后，首次实验以了解这些衔接蛋白及其HSP 100伴侣蛋白在B的蛋白质质量控制网络中的体内作用。subtilis介绍。