Proteomics of bile acid signals

本研究试图通过蛋白质组学的方法，建立胆汁酸信号转导的分析方法，阐明胆汁酸在脑内的功能。鹅去氧胆酸腺苷酸对生长激素的亲和标记分析结果表明，鹅去氧胆酸与生长激素受体结合位点不同的区域结合。接下来，我们开发了一种分析方法的磷酸化蛋白质使用特定的衍生化和双伪中性损失提取nanoLC/ESI-MS/MS。我们发现它的有用性，在复杂的蛋白质混合物中的磷酸化蛋白质的鉴定和磷酸化位点的鉴定。此外，我们通过使用鹅去氧胆酸盐固定化可切割亲和凝胶提取肝细胞中的鹅去氧胆酸盐结合蛋白，并鉴定了肝细胞中的胆汁酸结合蛋白二氢二醇脱氢酶和过氧化物氧还蛋白1。

According to proteomics approach, we tried to develop an analytical method for bile acid signal transduction and clarify the function of bile acids in the brain. The results by the affinity labeling analysis of growth hormone using chenodeoxycholyl adenylate showed that chenodeoxycholate binds to the region different from the growth hormone receptor binding site. Next, we developed an analytical method of phosphorylated proteins using the specific derivatization and the double pseudo-neutral loss extraction on nanoLC/ESI-MS/MS. We found its usefulness for identification of phosphorylated proteins in complex protein mixtures and identification of phosphorylated sites. In addition, we extracted chenodeoxycholate-binding proteins in hepatocytes by using the chenodeoxycholate-immobilized cleavable affinity gel, and identified dihydrodiol dehydrogenase, which is known as a bile acid-binding protein in the hepatocyte, and peroxiredoxin 1.