PROTEOMICS-BASED ANALYSIS OF BLOOD-BRAIN BARRIER TRANSPORT SYSTEM

在液相色谱法分离鉴定膜蛋白的基础上，建立了一种高灵敏度、高同步的膜蛋白定量方法。串联质谱（LC-MS/MS）与多路多重/选择性反应监测（MRM）模式。最重要的结果是，我们成功地建立了一个计算机选择标准，以找到合适的肽片段进行定量，并将在LC-MS/MS分析中获得高强度。利用该方法，我们定量测定了小鼠脑毛细血管内皮细胞即血脑屏障中的转运蛋白。此外，我们成功地阐明了转运蛋白在人血脑屏障中的定量表达。我们鉴定了11种可能在人血脑屏障中起转运作用的膜蛋白。这些发现将对今后的血脑屏障研究，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.