Proteomics analysis of plasma membrane from liver sinusoidal endothelial cells after partial hepatectomy by an improved two-dimensional electrophoresis
Proteomics analysis of plasma membrane from liver sinusoidal endothelial cells after partial hepatectomy by an improved two-dimensional electrophoresis
复制标题
改进的二维电泳对部分肝切除术后肝窦内皮细胞质膜进行蛋白质组学分析
DOI：
10.1007/s11010-010-0537-z
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发表时间：
2010-11-01
影响因子：
4.3
通讯作者：
Liang, Songping
中科院分区：
文献类型：
--
作者：
Li, Xuanwen;Xiong, Li;Liang, Songping
肝再生是一种血管生成相关的现象。为了确定肝再生过程中内皮细胞参与启动血管生成的关键质膜蛋白，采用体内膜密度微扰法对肝部分切除后72h的肝窦内皮细胞(LSEC)质膜蛋白进行了浓缩。用改进的二维16-BAC/SDS-PAGE对与假手术相比差异表达的膜蛋白进行定量，并用LC-MS/MS对其中的几种蛋白质进行鉴定，并用CICAT标记定量策略进一步证实了其中一些蛋白质。共鉴定出47种蛋白质，包括与血管生成或肝再生有关的已知和新蛋白质，如诱导型一氧化氮合酶、IV型胶原和整合素Beta3。我们的结果表明，膜密度微扰法和改进的双向电泳(2-DE)方法相结合是研究体内内皮功能障碍的有效方法。
Liver regeneration is an angiogenesis-associated phenomenon. To identify key plasma membrane (PM) proteins of endothelial cells involved in the initiation of angiogenesis during liver regeneration, the PM of liver sinusoidal endothelial cells (LSEC) at 72 h after partial hepatectomy was enriched by an established in vivo membrane density perturbation method. The differentially expressed membrane proteins compared to those from sham operation were quantified using an improved two-dimensional 16-BAC/SDS-PAGE and identified by LC-MS/MS. Several proteins were further confirmed by cICAT labeling quantitative strategy. A total of 47 proteins were identified including known and novel proteins involved in angiogenesis or liver regeneration, such as inducible nitric oxide synthase, type IV collagen, and integrin beta3. Our results indicated that the combination of the membrane density perturbation strategy and the improved two-dimensional electrophoresis (2-DE) method are useful for investigating the endothelial dysfunctions in vivo.