Proteomic analysis of high-density lipoprotein
Proteomic analysis of high-density lipoprotein
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DOI：
10.1002/pmic.200500191
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发表时间：
2006-01-01
期刊：
影响因子：
3.4
通讯作者：
Meijers, JCM
中科院分区：
文献类型：
--
作者：
Rezaee, F;Casetta, B;Meijers, JCM
血浆脂蛋白，如高密度脂蛋白（HDL），可以作为广泛的蛋白质的载体，参与过程，如脂质代谢，血栓形成，炎症和动脉粥样硬化。HDL相关蛋白的鉴定对于在分子水平上理解这些过程至关重要。在这项研究中，结合蛋白质组学方法，包括1-DE和2-DE MALDI-TOF，同位素编码的亲和标签和蛋白质印迹分析，以确定与人类HDL相关的蛋白质。为了最大限度地减少分离过程中HDL相关蛋白的潜在损失，应用一步超离心技术，并通过浊度法高效凝胶色谱法和Western印迹分析确认纯化HDL的质量。MS分析显示存在56种HDL相关蛋白，包括所有已知的载脂蛋白和脂质转运蛋白。此外，还发现了与止血、血栓形成、免疫和补体系统有关的蛋白质，以及生长因子、受体、血管相关蛋白等多种蛋白质。因此，我们的方法鉴定了大量与HDL相关的蛋白质。蛋白质组学技术的结合被证明是一个强大的和全面的工具，用于识别蛋白质的HDL。
Plasma lipoproteins, such as high-density lipoprotein (HDL), can serve as carriers for a wide range of proteins that are involved in processes such as lipid metabolism, thrombosis, inflammation and atherosclerosis. The identification of HDL-associated proteins is essential with regards to understanding these processes at the molecular level. In this study, a combination of proteomic approaches including 1-DE and 2-DE MALDI-TOF, isotope-coded affinity tag and Western blot analysis were employed to identify proteins associated with human HDL. To minimize potential losses of HDL-associated proteins during isolation, a one-step ultracentrifugation technique was applied and the quality of purified HDL was confirmed by nephelometry highperformance gel chromatography, and Western blot analysis. MS analysis revealed the presence of 56 HDL-associated proteins including all known apolipoproteins and lipid transport proteins. Furthermore, proteins involved in hemostasis and thrombosis, the immune and complement system were found. In addition, growth factors, receptors, hormone-associated proteins and many other proteins were found to be associated with HDL. Our approach thus resulted in the identification of a large number of proteins associated with HDL. The combination of proteomic technologies proved to be a powerful and comprehensive tool for the identification of proteins on HDL.