Communication between the domains of the DnaK chaperone characterized by single molecule force spectroscopy

通过单分子力谱表征的 DnaK 伴侣域之间的通信

• 批准号: 170292651
• 负责人: Dr. Gabriel Zoldák
• 金额: --
• 依托单位: Lehrstuhl für Molekulare Biophysik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2010
• 资助国家: 德国
• 起止时间: 2009-12-31 至 2011-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/170292651?language=en
• 关键词: Communication; between; domains; DnaK; chaperone

DnaK is a Hsp70 chaperone of E. coli which consists of an N-terminal nucleotide binding domain (NBD) and a C-terminal substrate binding domain (SBD). NBD binds ATP or ADP and displays very weak ATPase activity. SBD recognizes hydrophobic stretches in the polypeptide chain of protein substrates. DnaK shows negative cooperativity between the binding of ATP to NBD and protein substrates to the remote SBD. This bidirectional allosteric communication is mediated through the conserved linker between the domains. Notably, the underlying physical principles of the communication pathways are still not know. Recent progress in atomic force microscopy (AFM) allows molecular forces to be measured at sub-pico-Newton resolution and at variable dimensions what could be particularly interesting for allosteric proteins. Using these techniques, my major goals will be to elucidate: (1) the interdomain dynamics of DnaK in different states; (2) the mechanics of the allosteric transitions upon nucleotide and substrate binding; and (3) the movements of DnaK subdomains induced by the concerted action of the co-chaperone DnaJ and the nucleotide exchange protein GrpE.

DNAK是大肠杆菌的Hsp70分子伴侣，由N端核苷酸结合区(NBD)和C端底物结合区(SBD)组成。NBD与ATP或ADP结合，表现出很弱的ATPase活性。SBD识别蛋白质底物多肽链中的疏水伸展。DNAK在ATP与NBD的结合和蛋白质底物与远端SBD的结合之间表现出负的协同作用。这种双向变构通讯是通过结构域之间的保守连接子来调节的。值得注意的是，通信路径的潜在物理原理仍然不清楚。原子力显微镜(AFM)的最新进展使得可以在皮牛顿以下的分辨率和可变的维度测量分子力，这对于变构蛋白质来说可能是特别有趣的。利用这些技术，我的主要目标将是阐明：(1)不同状态下DNAK的结构域间动力学；(2)核苷酸和底物结合时变构转换的机制；以及(3)共同伴侣DNAJ和核苷酸交换蛋白GRPE协同作用引起的DNAK亚域的移动。

项目成果

Force as a single molecule probe of multidimensional protein energy landscapes.

• DOI: 10.1016/j.sbi.2012.11.007
• 发表时间: 2013-02
• 期刊: Current opinion in structural biology
• 影响因子: 6.8
• 作者: G. Žoldák;M. Rief