A Holistic Approach to Understanding Small Heat Shock Protein Mechanism

了解小热激蛋白机制的整体方法

• 批准号: 10615075
• 负责人: Rachel E Klevit
• 金额: $ 40.63万
• 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 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10615075
• 关键词: Age related macular degeneration; Aging; Belief; Binding; Blindness; Cataract; Cell Aging; Cells; Cellular Stress; Chimera organism; Chronic stress; Client; Complex; Comprehension; Cornea; Crystalline Lens; Crystallins; Data; Deuterium; Development; Diabetic Retinopathy; Disease; Exposure to; Eye; Failure; Genetic; Genetic Transcription; Goals; Grain; Grant; HSPB1 gene; Health; Heat shock proteins; Human; Hydrogen; Hypoxia; Individual; Injury; Ischemia; Knowledge; Link; Maintenance; Mass Spectrum Analysis; Modeling; Modification; Molecular; Molecular Chaperones; Molecular Conformation; Mutation; N-terminal; Neurodegenerative Disorders; Ocular Pathology; Pathology; Phosphorylation; Physiological; Positioning Attribute; Proteins; Resolution; Rest; Retina; Smoking; Specificity; Stress; Structure; Surface; Techniques; Testing; Tissues; Ultraviolet Rays; coping; crosslink; first responder; holistic approach; lens; lens transparency; novel; programs; protein aggregation; protein function; protein misfolding; protein structure; response; tau Proteins; tau aggregation; unnatural amino acids

ABSTRACT/SUMMARY Cells have numerous strategies to cope with the consequences of stresses that cause protein misfolding and aggregation and lead 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. Ocular tissues are subjected to stresses such as exposure to UV light, smoking, hypoxia, and ischemia. sHSP function is linked to three of the most prevalent ocular pathologies leading to blindness worldwide: cataract, age-related macular degeneration, and diabetic retinopathy which together account for 65% of world blindness. In lens, sHSPs perform the critical task of maintaining lens transparency and failure to do so is directly linked to cataract. sHSPs are expressed constitutively in retinal cells and are upregulated following injury or stress. Mechanisms used by sHSPs to delay the onset of aggregation of proteins remain enigmatic due to technical challenges posed by sHSPs and the aggregate-prone proteins they protect. Recent developments in the study of disordered proteins and breakthroughs made during the previous period of this long-standing project promise to overcome this critical barrier to mechanistic understanding. Techniques such as NMR, hydrogen-deuterium exchange/mass spectrometry, and covalent cross-linking/mass spectrometry can provide fine-grained information regarding disordered regions of sHSPs that have largely gone uncharacterized but are known to be essential for sHSP activity. The goal of this renewal application is to develop a holistic (“characterized by comprehension of the parts of something as intimately interconnected and explicable only by reference to the whole”) understanding of sHSP structure and function. The effects of stress conditions, modifications, and mutations will be assessed and interpreted in the context of the resultant novel models.

摘要/总结 细胞有多种策略来应对导致蛋白质产生的应激后果 错误折叠和聚集并导致斑块、原纤维和其他聚集物的形成 细胞衰老、白内障和神经退行性疾病中遇到的物种。蛋白质 被称为小热休克蛋白的伴侣是细胞的第一反应者，因此 维持细胞健康的关键。眼组织承受压力，例如 暴露于紫外线、吸烟、缺氧和缺血。 sHSP 功能与其中的三个相关 全球最常见的导致失明的眼部疾病：白内障、年龄相关性黄斑 变性和糖尿病性视网膜病变合计占世界失明率的 65%。在 晶状体中，sHSP 执行维持晶状体透明度的关键任务，如果未能做到这一点， 与白内障有直接关系。 sHSP 在视网膜细胞中组成型表达， 受伤或压力后上调。 sHSP 用于延迟发作的机制 由于 sHSP 和 它们保护易于聚集的蛋白质。无序蛋白质研究的最新进展 以及这个长期项目前期取得的突破 克服机械理解的这一关键障碍。核磁共振等技术， 氢-氘交换/质谱和共价交联/质量 光谱测定可以提供有关 sHSP 无序区域的细粒度信息， 大部分尚未被表征，但已知它们对于 sHSP 活性至关重要。目标是 这个更新应用程序是为了开发一个整体（“其特征是理解各个部分 某些事物是紧密相连的，并且只能通过整体来解释”） 了解 sHSP 结构和功能。压力条件、修改的影响， 突变将在由此产生的新模型的背景下进行评估和解释。