A Holistic Approach to Understanding Small Heat Shock Protein Mechanism

摘要/摘要 细胞有许多策略来应对导致蛋白质的压力的后果。 错误折叠和聚集，并导致斑块、纤维和其他聚集的形成 在衰老细胞、白内障和神经退行性疾病中遇到的物种。这种蛋白质 被称为小热休克蛋白的伴侣蛋白是细胞的第一反应者，因此 维持细胞健康的关键。眼组织会受到压力，如 暴露在紫外线下，吸烟，缺氧和缺血。SHSP函数链接到三个 全球最常见的致盲眼病：白内障、老年性黄斑 变性和糖尿病视网膜病变加起来占世界失明的65%。在……里面 透镜，sHSP执行维持透镜透明度的关键任务，而未能做到这一点是 与白内障有直接联系。SHSPs在视网膜细胞中有结构性表达，并且 在受伤或压力后表达上调。热休克蛋白用来延迟发病的机制 蛋白质的聚集仍然是个谜，因为sHSP和 它们保护的易于聚集的蛋白质。无序蛋白质研究的新进展 以及在这一长期项目的前一段时间取得的突破承诺 克服这一机械理解的关键障碍。核磁共振等技术 氢-氚交换/质谱学和共价交联/质量法 光谱分析可以提供关于sHSPs无序区域的细粒度信息 在很大程度上没有确定其特征，但已知对SHSP活动是必不可少的。的目标是 这个续订申请是为了开发一个整体的(特征是理解的部分 只有通过参照整体才能紧密联系和解释的东西“) 了解SHSP的结构和功能。压力条件的影响，修改， 突变将在所产生的新模型的背景下进行评估和解释。

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.