Protein Supported Lipid Bilayers as a Mimic of Native Biological Membranes

描述(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。许多使人类衰弱的疾病与蛋白质聚集体在组织中的沉积有关，这些蛋白质聚集体被称为淀粉样纤维。该项目的主要目标是表征淀粉样原纤维形成早期阶段的中间产物。本研究将以核糖核酸酶Sa作为模型系统。最近，肖尔茨实验室研究了RNaseSa和两个变异体在TFe存在下形成纤维的倾向以及作为pH的函数。在初步研究中，我们开发了一些条件，在这些条件下，我们已经在形成淀粉样纤维的溶液中鉴定和分离了几个小分子质量的RNaseSa寡聚体。我将通过近紫外光-环糊精、荧光、有限的蛋白质分解，可能通过核磁共振和氢交换质谱学来表征纤维形成的二级和三级结构。总的目标是从分子上了解淀粉样蛋白纤维形成的早期事件，从而有希望地设计小分子或治疗策略，以帮助防止这些蛋白质错误折叠疾病造成的毁灭性后果。

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.