Lipoproteomics II: Mapping of proteins in high-density lipoprotein using two-dimensional gel electrophoresis and mass spectrometry
Lipoproteomics II: Mapping of proteins in high-density lipoprotein using two-dimensional gel electrophoresis and mass spectrometry
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DOI：
10.1002/pmic.200401010
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发表时间：
2005-04-01
期刊：
影响因子：
3.4
通讯作者：
Lindahl, M
中科院分区：
文献类型：
--
作者：
Karlsson, H;Leanderson, P;Lindahl, M
高密度脂蛋白(HDLE)是血浆中含量最丰富的脂蛋白颗粒，是动脉粥样硬化的负性危险因素。通过使用蛋白质组学方法，有可能获得关于其蛋白质含量和蛋白质修饰的详细信息，这可能给出关于高密度脂蛋白的生理作用的新信息。本研究采用两步不连续密度梯度超速离心法分离HDL2和HDL3两个亚组分，用双向凝胶电泳法分离蛋白质，并用基质辅助激光解吸/电离飞行时间质谱仪进行多肽指纹图谱鉴定。在高密度脂蛋白中鉴定的蛋白质是：主要的载脂蛋白A-I为六种亚型，其中四种是糖基化模式，一种是保留的前肽，载脂蛋白A-II，载脂蛋白A-IV，载脂蛋白C-I，载脂蛋白C-II，载脂蛋白C-III(两种亚型)，载脂蛋白E(五种亚型)，最近发现的载脂蛋白M(两种亚型)，血清淀粉样蛋白A(两种亚型)和血清淀粉样蛋白A-IV(六种亚型)。此外，还首次在高密度脂蛋白中发现了α-1-抗胰蛋白酶。此外，唾液α-淀粉酶在HDL2中被鉴定为两种异构体，而在HDL3中被鉴定为载脂蛋白L和糖基化的载脂蛋白A-II。除了证实不同载脂蛋白的存在外，本研究还指出了糖基化载脂蛋白A-I和载脂蛋白A-II的新模式。此外，这项研究揭示了高密度脂蛋白中的新蛋白质：α-1-抗胰蛋白酶和唾液α-淀粉酶。对这些蛋白质的进一步研究可能会对高密度脂蛋白在冠状动脉疾病中的功能作用提供新的见解。
High-density lipoprotein (HDL) is the most abundant lipoprotein particle in the plasma and a negative risk factor of atherosclerosis. By using a proteomic approach it is possible to obtain detailed information about its protein content and protein modifications that may give new information about the physiological roles of HDL. In this study the two subfractions; HDL2 and HDL3, were isolated by two-step discontinuous density-gradient ultracentrifugation and the proteins were separated with two-dimensional gel electrophoresis and identified with peptide mass fingerprinting, using matrix-assisted laser desorption/ionisation time of flight mass spectrometry. Identified proteins in HDL were: the dominating apo A-I as six isoforms, four of them with a glycosylation pattern and one of them with retained propeptide, apolipoprotein (apo) A-II, apo A-IV, apo C-I, apo C-II, apo C-III (two isoforms), apo E (five isoforms), the recently discovered apo M (two isoforms), serum amyloid A (two isoforms) and serum amyloid A-IV (six isoforms). Furthermore, alpha-1-antitrypsin was identified in HDL for the first time. Additionally, salivary alpha-amylase was identified as two isoforms in HDL2, and apo L and a glycosylated apo A-II were identified in HDL3. Besides confirming the presence of different apolipoproteins, this study indicates new patterns of glycosylated apo A-I and apo A-II. Furthermore, the study reveals new proteins in HDL; alpha-1-antitrypsin and salivary alpha-amylase. Further investigations about these proteins may give new insight into the functional role of HDL in coronary artery diseases.