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采用RPLC-SDS-PAGE与纳升毛细管液相色谱-串联质谱联用技术对国际人类蛋白质组织(HUPO)提供的法国人肝脏蛋白质组表达谱的构建及其理化性质的统计分析,鉴定了2552条肽段,归结于402种蛋白质和240个蛋白质簇。蛋白质的pI分布范围为:4.29～11.57,分子量范围为:8,593～3,816,218Da,其中大于100kDa的蛋白质占到所鉴定蛋白质的10%,二者皆高于两维凝胶电泳的等电点和分子量范围(一般等电点3～10;分子量10～100kDa)。另外,蛋白质组中不同蛋白质的疏水性分布显示大部分蛋白质为可溶性蛋白质,其中疏水性蛋白质占15%(GRAVY>0的蛋白质)。实验结果表明该技术路线兼顾了反相色谱分离度高和SAS-PAGE兼容性好的优点,可用于复杂组织或细胞中蛋白质组表达谱的构建或比较蛋白质组的研究,为蛋白质组学的方法学研究奠定了基础。
RP-SDS-PAGE and nanoliter capillary liquid chromatography-tandem mass spectrometry (LC-MS / MS) techniques were used to characterize the expression profiles of the liver proteins of the French human liver provided by the International Human Proteins Organization (HUPO) and their physical and chemical properties. 2552 Peptides, due to 402 proteins and 240 protein clusters. The pI distribution ranged from 4.29 to 11.57 and the molecular weight ranged from 8,593 to 3,816,218 Da. Proteins larger than 100 kDa accounted for 10% of the identified proteins, both of which were higher than the isoelectric point and molecular weight of two-dimensional gel electrophoresis Range (usually isoelectric point 3 ~ 10; molecular weight 10 ~ 100kDa). In addition, the hydrophobic distribution of different proteins in the proteome revealed that most of the proteins were soluble, with 15% of hydrophobic proteins (GRAVY> 0). The experimental results show that this technique has the advantages of high resolution of reversed-phase chromatography and good compatibility of SAS-PAGE. It can be used to construct proteome expression profiles in complex tissues or cells or to compare proteomes. Proteomics methods Study laid the foundation for the study.