Research Institute of Pharmaceutical Sciences, Musashino University, Tokyo, Japan.
Biol Pharm Bull. 2012;35(9):1393-400. doi: 10.1248/bpb.b212006.
To study biochemical events in tissues and cells, we have developed a novel proteomics approach, FD-LC-MS/MS, which consists of fluorogenic derivatization (FD), LC separation and detection/quantification of proteins in a biological sample, followed by the isolation and tryptic digestion of target proteins, and then their identification using nano-HPLC-MS/MS. Fluorogenic reagents such as 7-chloro-N-[2-(dimethylamino)ethyl]-2,1,3-benzoxadiazole-4-sulphonamide (DAABD-Cl) were designed to have high sensitivity for HPLC-fluorescence and -MS/MS detection and reactivity for cysteine residues in proteins. This comprehensive differential proteomics approach was applied to several tissues, such as mouse liver, mouse brain, horse muscle, breast cancer cell lines, and mouse heart, in order to study fluctuations in protein levels in tissues and cells.
为了研究组织和细胞中的生化事件，我们开发了一种新的蛋白质组学方法 FD-LC-MS/MS，它包括荧光衍生化 (FD)、LC 分离以及对生物样本中蛋白质的检测/定量，随后是目标蛋白质的分离和胰酶消化，然后使用纳升 HPLC-MS/MS 对其进行鉴定。荧光衍生试剂，如 7-氯-N-[2-(二甲氨基)乙基]-2,1,3-苯并恶二唑-4-磺酰胺 (DAABD-Cl)，被设计用于具有高灵敏度的 HPLC-荧光和-MS/MS 检测以及对蛋白质中半胱氨酸残基的反应性。这种全面的差异蛋白质组学方法被应用于几种组织，如小鼠肝脏、小鼠大脑、马肌肉、乳腺癌细胞系和小鼠心脏，以研究组织和细胞中蛋白质水平的波动。
