Molecular aspects of copper binding to the prion protein

描述（由申请人提供）：本研究计划的目的是在分子水平上阐明朊蛋白（PrP）中铜和锌结合的机制和功能，以及这些相互作用与神经元维持和神经系统疾病的关系。朊病毒蛋白（PrP）的错误折叠形式导致了一类致命的神经退行性疾病，称为传染性海绵状脑病（tse），其中包括人类的疯牛病和克雅氏病。PrP是一种非常丰富的膜结合糖蛋白，存在于所有哺乳动物和鸟类中。虽然最初认识到它在神经退行性疾病中的作用，但现在清楚的是，PrP对于维持神经元功能和调节细胞凋亡至关重要。PrP在对PrP活性至关重要的部位吸收铜和锌。在上一个资助期内，PI的实验室表征了这些金属离子位点的结构和热力学特征，并确定了遗传性朊病毒疾病中某些突变与发病年龄之间的显著联系。下一个资助期将以这些发现为基础，利用磁共振和细胞研究进行以下项目：1)确定全长、铜或锌占据的蛋白质的结构；2)评估金属离子如何影响体内裂解，涉及细胞凋亡调节；3)研究与遗传疾病相关的突变如何改变结构和细胞行为，以及预防疾病的突变。此外，项目三将应用新的合成策略来研究蛋白质糖基化的作用。预计这些研究将为理解PrP和铜在基础神经化学中的作用以及治疗神经退行性疾病和脑外伤的策略提供基本的新概念。

DESCRIPTION (provided by applicant): The objective of this research program is to elucidate, at the molecular level, the mechanism and function of copper and zinc binding in the prion protein (PrP) and the relationship of these interactions to neuron maintenance and neurological disease. A misfolded form of the prion protein (PrP) is responsible for a class of fatal neurodegenerative diseases termed the Transmissible Spongiform Encephalopathies (TSEs), which include mad cow disease and Creutzfeldt-Jakob disease in humans. PrP is a highly abundant, membrane-bound glycoprotein found in all mammals and avian species. Although originally recognized for its role in neurodegenerative disease, it is now clear that PrP is essential for sustaining neuron function and regulating apoptosis. PrP takes up both copper and zinc at sites are essential for PrP activity. Over the last funding period, the PI's laboratory characterized structural and thermodynamic features of these metal ion sites and also identified a remarkable link between certain mutations and onset age in inherited prion disease. The next funding period will build on these findings using both magnetic resonance and cellular studies with the following projects: 1) determine the structure of the full-length, copper or zinc occupied protein; 2) evaluate how metal ions influence in vivo cleavage, implicated in apoptosis regulation; and 3) examine how structure and cellular behavior is altered by mutations associated with inherited disease, and also by mutations that prevent disease. Moreover, project three will apply new synthetic strategies for investigating the role of protein glycosylation. It is anticipated that these studies will provide fundamental new concepts for understanding the role of both PrP and copper in basic neurochemistry, and strategies for treating neurodegenerative disease and brain trauma. PUBLIC HEALTH RELEVANCE: This research is aimed at evaluating proteins of the central nervous system that regulate copper and other micronutrients. These proteins are essential for neuron health and understanding their behavior will lead to new strategies for treating neurodegenerative diseases, such as Alzheimer's, and treating brain trauma.