多氢键网格富集材料用于AD相关蛋白动态磷酸化和糖基化的同步研究

Aberrant protein phosphorylation and sialylated glycosylation have been closely related to Alzheimer's disease (AD). Because of the inherent limitations in intermolecular interactions, the TiO2 based enrichment method for phosphopeptides and sialylated glycopeptides (SGPs) suffers from low selectivity and recovery. So far, simultaneous exploring of AD related comprehensive phosphoproteomics and sialylated glycoproteomics has not been realized. Taking advantage of the specificity, orientation, and responsiveness of multiple hydrogen bonds (H-bonds) network in nature, coupled to the tunability and conformational transition of responsive polymers, we propose here to combine the dynamic H-bonding network with responsive polymers into the field of enrichment materials to solve the bottleneck of post-translational modification (PTM) proteomics, that is the simultaneous enrichment of phosphopeptides and SGPs. Firstly, the H-bonds based dipeptide receptors toward phosphopeptides and SGPs will be designed and screened out in order to solve the contradictions among selectivity, strong affinity and tunability in PTM-proteomics. After grating the optimized dipeptide receptor onto silica microspheres, the enrichment methods will be optimized and established. Meanwhile, we will pay attention to the adsorption/desorption dynamics between target peptides and adsorbents. Finally, the established method combined with quantitative proteomic technique will be applied to AD samples to find regulated phosphorylation and glycosylation sites. The multiple H-bonding network based responsive polymer will open an avenue and provide novel insight for material design of PTM-proteomics.

异常的蛋白质磷酸化和唾液酸N-糖基化与阿尔茨海默病(AD)密切相关。现有基于TiO2的磷酸化肽(PP)和唾液酸糖肽(SGP)同时富集方法因分子间作用力的限制，存在选择性差和回收率低等问题，难以实现AD相关全面磷酸化组学和唾液酸糖基化组学的同时分析。本课题将多氢键体系的专一性、定向性、响应性以及响应性聚合物的可调性和构象转变等理念引入到富集材料的设计中，以解决翻译后修饰组学研究的瓶颈问题。通过设计和筛选基于二肽的PP和SGP多氢键受体，解决选择性、强结合和可调控之间的矛盾；将筛选出的二肽受体聚合到硅球上制备响应性富集材料，建立PP和SGP同时富集、顺序洗脱方法；注重目标肽段在富集材料上的吸附/脱附动力学研究；结合定量蛋白质组学技术，将所建立富集方法应用于AD病变相关蛋白调控性修饰位点的发现。基于多氢键网格的富集材料为翻译后修饰蛋白组学开辟了新方向、发展了新思路。

异常的蛋白质磷酸化和唾液酸N-糖基化与阿尔茨海默病(AD)密切相关。现有基于TiO2的磷酸化肽(PP)和唾液酸糖肽(SGP)同时富集方法因分子间作用力的限制，存在选择性差和回收率低等问题，难以实现AD相关全面磷酸化组学和唾液酸糖基化组学的同时分析。本课题将多氢键体系的专一性、定向性、响应性以及响应性聚合物的可调性和构象转变等理念引入到富集材料的设计中，以解决翻译后修饰组学研究的瓶颈问题。通过设计和筛选基于二肽的PP和SGP多氢键受体，解决选择性、强结合和可调控之间的矛盾；将筛选出的二肽受体聚合到硅球上制备响应性富集材料，建立PP和SGP同时富集、顺序洗脱方法；注重目标肽段在富集材料上的吸附/脱附动力学研究；结合定量蛋白质组学技术，将所建立富集方法应用于AD病变相关蛋白调控性修饰位点的发现。基于多氢键网格的富集材料为翻译后修饰蛋白组学开辟了新方向、发展了新思路。