School of Life Sciences, Beijing Institute of Technology, Beijing, P. R. China.
Proteomics. 2013 Aug;13(16):2455-68. doi: 10.1002/pmic.201300073. Epub 2013 Jul 17.
The proteomic profile of hypothalamus, a key organ of CNS, is explored here by employing two widely used MS techniques, i.e. HPLC/ESI-ion trap and HPLC/ESI-quadrupole-TOF MS. Strong cation exchange is used for the fractionation of peptides and protein search engine MASCOT is employed for data query. One hundred and thirty six proteins with 10 973 peptides were identified by HPLC/ESI-ion trap MS, while 140 proteins with 32 183 peptides were characterized by HPLC/ESI-quadrupole-TOF MS. Among the total 198 proteins identified in both experiments, 78 proteins were common in both sets of conditions. The rest of the 120 proteins were identified distinctly in both MS strategies, i.e. 58 unique proteins were found using the quadrupole-TOF while 62 were found with the HPLC/ESI-ion trap. Moreover, these proteins were classified into groups based on their functions performed in the body. Results presented here identified some important signal and cellular defense proteins inevitable for survival in stressed conditions. Additionally, it is also shown that any single MS strategy is not reliable for good results due to loss of data depending on sensitivity of the instrument used.
通过采用两种广泛使用的 MS 技术，即 HPLC/ESI-离子阱和 HPLC/ESI-四极杆-TOF MS，探索了中枢神经系统关键器官下丘脑的蛋白质组特征。强阳离子交换用于肽的分离，并且使用 Mascot 数据查询搜索引擎进行数据查询。通过 HPLC/ESI-离子阱 MS 鉴定了 136 种蛋白质，有 10973 种肽，而通过 HPLC/ESI-四极杆-TOF MS 鉴定了 140 种蛋白质，有 32183 种肽。在这两种实验中总共鉴定的 198 种蛋白质中，78 种蛋白质在两种条件下都是共同的。其余的 120 种蛋白质在两种 MS 策略中都有明显的区别，即使用四极杆-TOF 发现了 58 种独特的蛋白质，而使用 HPLC/ESI-离子阱发现了 62 种蛋白质。此外，这些蛋白质根据其在体内执行的功能进行了分类。这里呈现的结果确定了一些在应激条件下生存所必需的重要信号和细胞防御蛋白质。此外，还表明由于所使用仪器的灵敏度不同，任何单一的 MS 策略都不可靠，因为会丢失数据。
