表面等离子体共振-质谱联用研究人载脂蛋白与细胞膜之间的相互作用

Apolipoproteins (Apo) are proteins that bind lipids to form lipoproteins, and are very important components of plasma lipoproteins. They showed many critical functions in biosystem, e.g., they can serve as enzyme cofactors, receptor ligands that regulate the metabolism of lipoproteins. More and more researches indicated that apolipoproteins have very close relationship with many diseases. Many physiological functions of apolipoproteins are related to its interactions with phospholipid membrane, thus the study of the interaction between apolipoproteins and phospholipids is of great significance. Surface plasmon resonance (SPR) is a sensitive surface analysis technique, which has been widely applied in the field involving interactions. The online coupling of SPR to mass spectrometry (MS) could facilitate the identification and quantitation of the molecules involved in the interaction and resulted products. Therefore, this project will develop hyphenated technique that couples SPR to ambient MS, then some novel sensor chips modified with cell membrane were used in this technique to investigate the interaction of human apolipoproteins with cell membrane, aiming to reveal their interaction mechanisms. The results will provide an effective SPE-MS method for interaction study and to obtain some important data for understanding the insight into the relationship between the interactions and some human diseases.

载脂蛋白(Apo)可与脂类结合形成脂蛋白，是构成血浆脂蛋白的重要组分，具有多种生物功能。研究表明，Apo与诸多人类疾病密切相关。由于Apo在体内的许多生理功能都和其与磷脂膜之间的相互作用有关，因此研究Apo与细胞膜之间的相互作用对于理解生命过程和疾病发生机制具有重要的价值。表面等离子体共振(SPR)技术在相互作用研究领域应用广泛，其与质谱(MS)的在线联用能够实现相互作用分子及其产物的实时、快速和原位检测，但技术尚欠成熟。本项目拟在前期工作的基础上，发展和完善SPR与敞开式离子化MS的联用方法，进而研究人Apo与细胞膜之间的相互作用。选取代表性的Apo A1, Apo B, Apo E作为目标分析物，将细胞膜修饰在芯片金膜表面，以研究各Apo与其相互作用情况，鉴定相互作用的产物，从而深入研究人Apo与细胞膜之间相互作用的机制，为揭示载脂蛋白与人类疾病之间的关系提供有效的研究方法和科学数据。

载脂蛋白(Apo)可与脂类结合形成脂蛋白，是构成血浆脂蛋白的重要组分，具有多种生物功能，且Apo与诸多人类疾病密切相关。由于Apo在体内的许多生理功能都和其与磷脂膜之间的相互作用有关，因此研究Apo与细胞膜之间的相互作用对于理解生命过程和疾病发生机制具有重要的价值。本项目拓展了表面等离子体共振(SPR)技术在相互作用研究领域的应用，发展和完善了SPR与敞开式离子化MS的联用方法。选取代表性的Apo A1, Apo B, Apo E作为目标分析物，将细胞膜修饰在芯片金膜表面，进而研究了人Apo与细胞膜之间的相互作用。采用外泌体修饰了芯片，深入研究了细胞外泌体与小分子之间的相互作用的机制。项目的完成为SPR提供了新的检测方法，也为生命科学中分子相互作用和药物筛选提供了有效的研究方法和科学数据。.项目成果共发表SCI论文11篇，为一部英文专著撰写了一章；培养博士研究生3名，博士后一名。基本完成了原计划的研究任务。