Mechanisms of Activation for Human Small Heat Shock Proteins: An Integrated Approach

人类小热休克蛋白的激活机制：综合方法

• 批准号: 9304219
• 负责人: Rachel E Klevit
• 金额: $ 61.53万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9304219
• 关键词: Affect; Affinity; Age related macular degeneration; Binding; Biological Assay; Blindness; Brain Hypoxia-Ischemia; Cataract; Cell Aging; Cells; Cellular Stress; Charge; Chronic; Client; Complex; Cornea; Crystalline Lens; Detection; Diabetic Retinopathy; Dimerization; Disease; Emerging Technologies; Equilibrium; Exhibits; Eye; Fluorescence Resonance Energy Transfer; Functional disorder; Goals; HSPB1 gene; Health; Heat shock proteins; Heat-Shock Response; Histidine; Human; Inherited; Ions; Knowledge; Lead; Link; Maintenance; Mammals; Mass Spectrum Analysis; Metal Ion Binding; Metals; Modeling; Molecular; Molecular Chaperones; Mutation; Names; National Eye Institute; Negative Staining; Neurodegenerative Disorders; Oxidative Stress; Pathology; Physiological; Play; Property; Proteins; Refractory; Regulation; Resolution; Retina; Role; Stress; Structural Models; Structure; Temperature; Testing; Tissues; UV Radiation Exposure; Work; Zinc; base; dimer; disease phenotype; divalent metal; dynamic system; emergency service responder; experience; experimental study; insight; lens; monomer; protein aggregation; protein function; protein misfolding; protein structure; public health relevance; response; solid state nuclear magnetic resonance; structural biology

 DESCRIPTION (provided by applicant): Cells have numerous strategies to cope with the consequences of stresses that cause protein misfolding and aggregation, leading to formation of plaques, fibrils, and other aggregated species encountered in aging cells, cataract, and neurodegenerative diseases. The protein chaperones known as small heat shock proteins are the cell's first responders and are therefore key to maintenance of cellular health. Despite their name, human small heat shock proteins (sHSP) are rarely called up to respond to the stress of elevated temperature, as humans have effective and tight regulation of temperature. Nevertheless, much of the current knowledge of sHSPs has been obtained under elevated temperature conditions where they are highly active. In this project, we seek to understand the molecular mechanisms by which human sHSPs respond to physiologically relevant stresses and how they maintain cellular proteins in soluble forms. Of relevance to the National Eye Institute, the sHSPs to be focused on are highly expressed in ocular tissues (lens, retina, cornea) as well as in other tissues. Dysfunction and aberrant expression of HSPB1 and HSPB5 are associated with ocular diseases including cataract, diabetic retinopathy, and age-related macular degeneration that together account for 65% of blindness worldwide. Perhaps linked to their particular functional niche in which they must respond to small changes in cellular conditions such as pH and metal ion concentrations, human sHSPs have evolved properties that are unique in the protein world. They exist as large (> 100 kDa) polydisperse and dynamic assemblies that defy conventional structural biology approaches. We will apply emerging technologies capable of providing molecular and possible atomic-level information about heterogeneous and dynamic systems. Structural information obtained from solution- and solid-state NMR, negative-stain and cryo-EM, and native mass spectrometry will be integrated with dynamic information from FRET-based subunit dynamics and NMR and with functional information from activity assays and client-binding experiments to define the mechanism(s) by which HSPB5 and HSPB1 are activated by conditions brought about by stresses relevant to tissues of the eye such as hypoxia, ischemia, and UV exposure. Both wild-type sHSPs and forms carrying inherited disease mutations will be investigated.

 描述（由适用提供）：细胞具有许多策略来应对引起蛋白质错误折叠和聚集的应力的后果，从而导致在衰老细胞，白内障和神经退行性疾病中遇到的斑块，原纤维和其他聚集物质形成。蛋白质伴侣称为小热激蛋白是细胞的第一个反应者，因此是维持细胞健康的关键。尽管人类的名称，但很少有人要求人类的小热冲击蛋白（SHSP）应对温度升高的压力，因为人类对温度有效且严格的调节。然而，在高度活跃的温度条件下，SHSP的许多当前知识都是获得了的。在这个项目中，我们试图了解人类SHSP响应物理相关的应力以及它们如何以固体形式维持细胞蛋白的分子机制。与国家眼科研究所相关的是，要关注的SHSP在眼组织（镜头，视网膜，角膜）以及其他组织中高度表达。 HSPB1和HSPB5的功能障碍和异常表达与包括白内障，糖尿病性视网膜病和与年龄相关的黄斑变性在内的眼部疾病有关，占全球失明的65％。也许与他们的特定功能性利基市场有关，在这些小众市场中，他们必须对细胞条件（例如pH和金属离子浓度）的微小变化做出反应，人类SHSP的发展性能在蛋白质世界中是独特的。它们以大型（> 100 kDa）的多分散和动态组件的形式存在，这些组合无需传统的结构生物学方法。我们将应用能够提供有关异质和动态系统的分子和可能的原子级信息的新兴技术。 Structural information obtained from solution- and solid-state NMR, negative-stain and cryo-EM, and native mass spectrometry will be integrated with dynamic information from FRET-based subunit dynamics and NMR and with functional information from activity assays and client-binding experiments to define the mechanism(s) by which HSPB5 and HSPB1 are activated by conditions bring about by stresses relevant to tissues of the eye such as hypoxia,缺血和紫外线暴露。将研究野生型SHSP和携带遗传性疾病突变的形式。