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.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 分子伴侣通过保护未折叠和错误折叠的蛋白质直到它们可以折叠或组装成复合物，然后帮助促进成功的折叠和组装，在细胞维持和应对压力中发挥重要作用。它们还与降解机制合作，以管理细胞的“质量控制”，并防止潜在有害的错误折叠产物的积累。伴侣共同作用形成一个复杂的蛋白质稳定网络，对细胞生存至关重要。伴侣蛋白的普遍存在的Hsp 70蛋白家族防止未折叠蛋白中的不正确相互作用，所述不正确相互作用可通过由ATP水解调节的客户蛋白的结合和释放的循环导致错误折叠或聚集，并进一步由辅伴侣蛋白如核苷酸交换因子（NEF）和Hsp 40调节。理解Hsp 70的细胞作用需要深入阐明Hsp 70变构机制和与辅助分子伴侣的相互作用;这对于开发基于Hsp 70分子伴侣调节的治疗策略至关重要。我们在ACERT设施的工作目标是获得关键信息，以全面描述E。coli Hsp 70，DnaK，在其整个功能性变构循环中以及在与辅分子伴侣相互作用时。我们将确定战略性地位于整个蛋白质中的各种自旋对之间的距离分布。这些距离分布将提供有关DnaK在各种核苷酸或底物或共分子伴侣结合状态下的结构的信息。此外，我们将使用双电子-电子共振（DEER）实验，以确定这些合奏稀疏填充的构象。这些配置文件，结合顺磁弛豫增强核磁共振数据正在收集的U。质量将使我们能够绘制出热休克蛋白70的变构景观。