β淀粉样前体蛋白的电化学传感新方法与相关分子机制研究

β淀粉样前体蛋白(APP)的代谢过程与淀粉样多肽(Aβ)的产生及阿尔茨海默病(AD)的发展有着密切的关联。因此开发有效的APP相关分子的检测方法，对AD发病机理的阐释及AD的诊断治疗至关重要，但目前此方面的研究还十分缺乏。本项目拟基于特异性生物偶联反应系统构建APP相关分子的新型电化学传感方法，并将其应用于APP相关分子生物学过程的考察及实际样品的检测，主要研究内容包括：①基于氨基酸特异性生物偶联反应构建APPβ分泌酶(BACE1)的比率型电化学检测方法；②基于磷酸及邻二醇基团的特异性识别反应构建APP多肽片段磷酸化及糖基化过程的检测方法；③根据构建的电化学传感体系，考察APP氨基酸突变及后修饰对BACE1酶切反应的影响；④通过实际样品的检测，探索与AD病理过程相关的APP分子机制及重要生物学参数。项目的执行将为AD的病理研究、诊断及药物开发提供有效的技术手段与理论参考。

The metabolic process of APP (β-amyloid precursor protein) is highly related to the generation Aβ (amyloid-β peptide) and the progress of AD (Alzheimer's disease). Thus, the development of effective sensing technique for monitoring APP relevant biomolecules is particularly important for elucidating the pathological mechanism of AD and also its diagnosis and treatment. However, the relevant studies have seldom been conducted up to now. In this project, we propose firstly to develope novel electrochemical sensing methodology aiming to APP, and then apply it to investigate the molecular biological process of APP as well as the real sample related to AD patient, which contains: (1) constructing ratiometric electrochemical sensor for BACE1 (β-site APP cleaving enzyme 1) based on the specific bioconjugation reaction for target amino acids; (2) establishing electrochemical sensor for the phosphorylation or glycosylation of APP fragment by exploiting the speccific reaction of phosphate group or vicinal diol; (3) investigating the relation between the amino acid mutation, phosphorylation, glycosylation of APP with the reaction behavior of BACE1 using the developed electrochemical sensors; (4) detecting the reactitities of BACE1, phosphorylase, and glycosylase in simulated and real samples and exploring the vital biological parameters and the molecular biology of APP correlated to AD. The implement of the project will provide the theoretical basis and technical support for studying the pathology of AD as well as diagnosis and treatment of AD.