THE ALLOSTERIC MECHANISM OF HSP70

HSP70 的变构机制

• 批准号: 8364103
• 负责人: LILA M GIERASCH
• 金额: $ 0.08万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8364103
• 关键词: ATP Hydrolysis; Binding; Cell Survival; Client; Complex; Data; Electrons; Escherichia coli; Funding; Goals; Grant; Guanine Nucleotide Exchange Factors; Heat-Shock Proteins 70; Lead; Maintenance; Maps; Molecular Chaperones; National Center for Research Resources; Nucleotides; Play; Principal Investigator; Protein Family; Proteins; Quality Control; Relaxation; Research; Research Infrastructure; Resources; Role; Source; Stress; Structure; Technology; Therapeutic; United States National Institutes of Health; Work; base; conformer; cost; prevent; protein misfolding; research study; response

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Molecular chaperones play essential roles in cellular maintenance and response to stress by protecting unfolded and misfolded proteins until they can be folded or assembled into complexes and then helping promote successful folding and assembly. They also partner with degradation machinery to manage cellular 'quality control' and prevent accumulation of potentially deleterious misfolded products. Chaperones work together to form a complex proteostasis network that is essential for cell survival. The ubiquitous Hsp70 protein family of chaperones prevents incorrect interactions in unfolded proteins that can lead to misfolding or aggregation by cycles of binding and release of client proteins regulated by ATP hydrolysis and further modulated by co-chaperones such as nucleotide exchange factors (NEFs) and Hsp40s. Understanding cellular roles of Hsp70s requires in-depth elucidation of the Hsp70 allosteric mechanism and interactions with co-chaperones; this is essential in order to develop therapeutic strategies based on modulation of Hsp70 chaperones. The goal of our work at the ACERT facility is to obtain crucial information for a full description of the structural ensembles of the E. coli Hsp70, DnaK, throughout its functional allosteric cycle and upon interaction with co-chaperones. We will determine distance profiles between various spin pairs strategically located throughout the protein. These distance profiles will provide information about the structures populated by DnaK in various nucleotide- or substrate- or co-chaperone-bound states. In addition, we will use double electron-electron resonance (DEER) experiments to identify sparsely populated conformers within these ensembles. These profiles, in conjunction with paramagnetic relaxation enhancement NMR data being collected at U. Mass will enable us to map out the allosteric landscape of an Hsp70.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 分子伴侣通过保护展开和错误折叠的蛋白质，直到可以将它们折叠或组装成复合物，然后帮助促进成功的折叠和组装，从而在细胞维持和对压力的反应中起着重要作用。他们还与降解机制合作，以管理蜂窝“质量控制”并防止潜在有害错误折叠产品的积累。伴侣共同形成一个复杂的蛋白抑制网络，该网络对于细胞存活至关重要。无处不在的HSP70蛋白质家族的伴侣蛋白家族阻止了未折叠的蛋白质中的不正确相互作用，这可能导致通过ATP水解调节的结合和释放的客户蛋白的循环折叠或聚集，该蛋白质受ATP水解的调节，并由共芯酮（如核苷酸交换因子（NEFS）和HSP40S）进一步调节。了解HSP70S的细胞作用需要深入阐明HSP70变构机制以及与副酮的相互作用。这对于基于HSP70伴侣的调节制定治疗策略至关重要。我们在ACERT设施的工作的目的是获得关键信息，以完整描述大肠杆菌HSP70，DNAK的结构集合，并在其功能变构循环中以及与共伴侣相互作用后。我们将在整个蛋白质中策略性地位于各种自旋对之间的距离曲线。这些距离曲线将提供有关DNAK在各种核苷酸或底物或副酮结合状态下人群的结构的信息。此外，我们将使用双电子电子共振（DEER）实验来识别这些团体中占量的构象异构体。这些轮廓以及在U.质量收集的顺磁性松弛增强NMR数据的结合，将使我们能够绘制出HSP70的变构景观。