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、生物技术和医学中一个极其重要的课题。许多使人衰弱的疾病与被称为淀粉样蛋白原纤维的蛋白质聚集体在组织中的沉积有关。这个项目的主要目标是在淀粉样纤维形成的早期阶段的中间体的特点。在本研究中，RNase Sa将被用作模型系统。最近，Scholtz实验室研究了RNase Sa和两种变体在TFE存在下形成原纤维的倾向，并作为pH值的函数。在初步研究中，我们已经开发了条件，在这些条件下，我们已经确定并分离了几种小分子量的RNase Sa低聚物，这些低聚物在形成淀粉样蛋白原纤维的过程中的溶液中。我将通过近紫外CD、荧光、有限蛋白水解、可能的核磁共振和氢交换质谱来表征原纤维形成的二级和三级结构。总体目标是从分子水平上了解淀粉样蛋白原纤维形成的早期事件，以期设计出小分子或治疗策略，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.