A Holistic Approach to Understanding Small Heat Shock Protein Mechanism

摘要/总结 细胞有许多策略来科普压力的后果，这些压力导致蛋白质 错误折叠和聚集，并导致形成斑块，原纤维，和其他聚集 在老化细胞、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分子伴侣是细胞的第一反应者，因此 维持细胞健康的关键。眼组织受到应力，例如 暴露于紫外线、吸烟、缺氧和局部缺血。sHSP功能与三种 全球最常见的致盲眼病：白内障、年龄相关性黄斑 视网膜变性和糖尿病性视网膜病变，它们共同占世界失明的65%。在 透镜、sHSP执行维持透镜透明度的关键任务，并且不能这样做是 与白内障直接相关。sHSP在视网膜细胞中组成型表达， 在损伤或应激后上调。sHSPs用于延迟发病的机制 由于sHSPs带来的技术挑战，蛋白质的聚集仍然是个谜， 它们所保护的易于聚集的蛋白质。无序蛋白质的研究进展 在这个长期项目的前期取得的突破承诺， 克服这一关键障碍，以机械的理解。诸如NMR的技术， 氢-氘交换/质谱和共价交联/质谱 光谱法可以提供关于sHSP的无序区域的细粒度信息， 在很大程度上没有得到表征，但已知对sHSP活性至关重要。的目标 这种更新应用程序是开发一个整体（“特征在于理解的部分， 只有参照整体才能解释的事物”） 了解sHSP的结构和功能。应力条件的影响，修改， 并且突变将在所得到的新模型的背景下被评估和解释。

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.