PROTEOMICS-BASED ANALYSIS OF BLOOD-BRAIN BARRIER TRANSPORT SYSTEM

在蛋白质消化液分离鉴定的基础上，我们建立了一种高灵敏的同时定量膜蛋白的方法。采用多重/选择性反应监测(MRM)模式的串联串联质谱仪(LC-MS/MS)。最重要的结果是，我们成功地建立了一种电子选择标准，以找到合适的多肽片段进行定量，并通过LC-MS/MS分析获得高强度的多肽片段。利用这种方法，我们对小鼠脑内毛细血管内皮细胞，即血脑屏障中的转运蛋白进行了定量。此外，我们成功地阐明了定量转运蛋白在人血脑屏障中的表达。我们鉴定了11种膜蛋白，它们可能是人血脑屏障中的转运蛋白。这些发现将对今后血脑屏障的研究，特别是对血脑屏障功能影响的认识产生重要影响。

We have developed a highly sensitive and simultaneous method to quantify membrane protein on the basis of separation and identification of protein digestsby liquid chromatography. linked tandem mass spectrometry (LC-MS/MS)with multiplexed multiple/selective reaction monitoring (MRM)mode. The most significant result is that we succeeded to developed an in silico selection criteria to find appropriate peptide fragment to be quantified which will give high intensity by LC-MS/MS analysis. By using this method, we have quantified transporter proteins in the brain capillary endothelial cells, i. e., blood-brain barrier, in mouse. Moreover, we succeeded to clarify quantitative transporter protein expression in the human blood-brain barrier. We identified 11 membrane proteins which may act as transporter in the human blood-brain barrier. These findings will give strong impact for the future blood-brain barrier research, especially for the understanding of effects on the blood-brain barrier function.