蛋白质乙酰化相关酶活性的动态传感研究及应用

Abstract: Aberrant protein acetylation is closely related with a variety of human diseases, especially cancer, therefore, developing real-time dynamic and highly sensitive analytical methods of protein acetylation related-enzymes are of great significance for cancer diagnosis/treatment and anti-cancer drug development. This project focuses on histone acetyltransferase and deacetylase based on the design and synthesis of novel functionalized acetylated peptide substrates and recognition unit. Moreover, the real-time variation of optical or electrochemical signals is explored by the property changes of the peptide substrate in acetylation process and the interaction between the peptide substrate and high-performance nanomaterials with many excellent properties. Then a series of novel real-time quantitative dynamic strategies on protein acetylation related-enzymes activity are developed by the specific binding of acetylation site and the self-characteristic of products generated by acetylation process. Furthermore the proposed methods have been continuously improved by a mixture of several realms, then further are applied to catch the abnormal signal of cancerous cells and screen inhibitors of acetylation related-enzymes. The project is to provide new directions and knowledge for the cancer diagnosis/treatment and anti-cancer drug development.

蛋白质乙酰化异常通常会诱发癌症等相关疾病的发生发展，发展蛋白质乙酰化相关酶活性的实时动态高敏分析方法对癌症诊疗和抗癌药物开发具有十分重要的意义。本项目拟将组蛋白乙酰化相关酶作为主要研究对象，设计、合成功能化多肽探针及识别单元，借助新型高性能纳米材料的优异特性，研究多肽底物乙酰化过程中自身性质变化及与高性能纳米材料间相互作用导致的光信号、电化学信号实时变化规律；利用乙酰化位点的特异性结合作用，联合乙酰化动态可逆修饰后产物的自身特质，进行多领域交叉研究，籍此开发乙酰化相关酶活性的实时高敏生物传感方法，实现活细胞内乙酰化相关酶活性的实时定量动态监测及癌变细胞的异常信号捕获，并用于筛选乙酰化相关酶的抑制剂。本项目以期为癌症诊疗和抗癌药物开发提供新方向和新知识。

研究组蛋白乙酰转移酶及其相关酶对于理解癌症成因和进展具有重要的意义。因此，发展新型的分析传感方法及平台，快速、高灵敏、多通道地动态监测癌症相关标志物及其发生发展过程，能够为癌症的预防、诊断、治疗以及预后执行提供方向。本项目以组蛋白乙酰转移酶及其相关酶（如乙酰胆碱酯酶、末端转移酶、核酸内切酶和外切酶等）作为主要研究对象，设计并合成了功能化多肽、抗体及适配体探针作为识别单元，借助新型高性能纳米材料（如石墨烯、金钯纳米材料、DNA纳米材料、类RNA配位聚合物等）的优异特性，研究多肽底物乙酰化过程中自身性质变化及与高性能纳米材料间相互作用导致的光信号、电化学信号实时变化规律；利用乙酰化位点的特异性结合作用，联合乙酰化动态可逆修饰后产物的自身特质，进行多领域交叉研究，开发了一系列乙酰化相关酶活性的实时高敏生物传感方法，并将此生物传感器用于小分子抑制剂的筛选。优异的灵敏度和选择性以及良好的重复性、重现性、稳定性和回收率说明了该传感器具有较好的应用前景，能够用于复杂体系中乙酰化相关肿瘤标志物的灵敏检测，对肿瘤的筛查、诊断、治疗和预防方面有着重要的指导意义。后续我们拟基于该项目进行拓展和深入研究，针对乙酰化/去乙酰化对细胞增殖分化的影响，寻找治疗恶性肿瘤最有效的手段，实现及早发现、诊断和治疗。