Protein Supported Lipid Bilayers as a Mimic of Native Biological Membranes

描述（由申请人提供）：蛋白质聚集是生物学、生物物理学、生物技术和医学中极其重要的课题。许多使人衰弱的人类疾病都与被称为淀粉样原纤维的蛋白质聚集体在组织中的沉积有关。该项目的主要目标是表征淀粉样蛋白纤维形成早期阶段的中间体。在本研究中，RNase Sa将作为模型系统。最近，Scholtz实验室研究了RNase Sa和两种变体在TFE存在和ph值作用下形成原纤维的倾向。在初步研究中，我们已经开发了条件，在形成淀粉样原纤维的过程中，我们已经确定并分离了几种RNase Sa的小分子量低聚物。我将通过近紫外- cd，荧光，有限的蛋白质水解，可能通过核磁共振和氢交换质谱来表征纤维形成的二级和三级结构。总体目标是对淀粉样蛋白纤维形成的早期事件进行分子理解，以期设计小分子或治疗策略，以帮助预防这些蛋白质错误折叠疾病的破坏性后果。

DESCRIPTION (provided by applicant): Protein aggregation is an extremely important subject in biology, biophysics, biotechnology, and medicine. A number of debilitating human diseases are associated with the deposition in tissue of protein aggregates known as amyloid fibrils. The primary goal of this project is to characterize the intermediates in the early stages of amyloid fibril formation. In this study, RNase Sa will be used as a model system. Recently, the Scholtz Lab has investigated the propensity of RNase Sa and two variants to form fibrils in the presence of TFE and as a function of pH. In preliminary studies, we have developed conditions in which we have identified and isolated several small molecular weight oligomers of RNase Sa in solutions that are in the process of forming amyloid fibrils. I will characterize the secondary and tertiary structure of fibril formation by near UV-CD, fluorescence, limited proteolysis, possibly by NMR, and hydrogen-exchange mass spectrometry. The overall goal is to reach a molecular understanding of the early events in amyloid fibril formation in an effort to hopefully design small molecules or treatment strategies to help prevent the devasting consequences of these protein misfolding diseases.