赤子爱胜蚓（Eisenia foetida）中抗肿瘤与纤溶酶原激酶活性蛋白质的分离与鉴定

采用SephadexG 75和HiPrep 1 6 / 6 0DEAE离子交换等方法从赤子爱胜蚓 (Eiseniafoetida)中提取到一组活性蛋白质成分 ,双向电泳分析证明它们为一组pI在 3 .0～ 4 .0之间的酸性蛋白质 ;利用体外K5 6 2、HeLa、SY5Y等肿瘤细胞抑制实验和纤维蛋白平板实验 ,跟踪测定活性 ,证明乙醇沉淀组分D2 (8)是既具肿瘤抑制 ,又具有激酶活性的蛋白质成分。同时应用非变性电泳对乙醇沉淀组分进行分离 ,并利用电洗脱、凝胶原位酶解和ESI MS等蛋白质组学方法 ,鉴定了其中 6种蛋白质的分子量、氨基酸组成、N末端序列和肽质量指纹图信息 ,其中条带 9与D2 (8)组分为同一种蛋白质 ;研究证明蚯蚓中含有既具抗肿瘤活性又具有激酶活性的蛋白质成分。采用的方法可适用于活性蛋白质成分的整体分离与鉴定

Isolation and Identification of Proteins with Anti-tumor and Fibrinolysogen Kinase Activities from Eisenia foetida

Proteins from Eisenia foetida possess many biological activities. A group of proteins precipitated by ethanol were isolated and purified by Sephadex G 75 and HiPrep 16/60 DEAE columns, then identified by one or two dimensional electrophoresis and mass spectrometry. 2D gel experiments displayed that the pI of proteins from Eisenia foetida were mainly from 3.0 to 4.0. Anti tumor and kinase activities were determined by in vitro experiments. The enthanol fraction D2(8) showed both of the activities. These ethanol precipitated proteins were identified further by native polyacrylamide electrophoresis, the protein spots were cut off from gels and digested by trypsin, the peptide mass fingerprints (PMFs) were determined by mass spectrometry. PMF, molecular weight , amino acid composition and N terminus of 6 proteins were characterized, and band 9 was identified as D2(8). The results suggested that there exist proteins in Eisenia foetida possessed both anti tumor and fibrinolysogen kinase activities. These methods can be used for identification of the natural bioactive proteins.