重组精氨酸脱亚胺酶的表达、纯化及其抗癌活性  

Expression,purification and anti-angiogenic activity analysis of arginine deiminase

作　　者：谭玲玲[1] 黄亮[1] 陈博[2] 

机构地区：[1]兰州大学生命科学学院,兰州730000 [2]复旦大学生命科学学院,上海200433

出　　处：《兰州大学学报（自然科学版）》2010年第2期60-65,共6页Journal of Lanzhou University(Natural Sciences)

基　　金：国家自然科学基金项目(30600040)

摘　　要：克隆了ADI并在大肠杆菌中表达重组蛋白,得到的ADI重组蛋白通过离子交换层析纯化并进一步复性.复性后的重组ADI蛋白经分析与天然ADI蛋白具有相近的分子量(大约46 kD)和相似的酶活性(38 U/mg),可以将L-精氨酸转换成瓜氨酸.在蛋白酶降解实验中,发现重组ADI具有比天然ADI更稳定的抗胰蛋白酶降解活性,重组ADI的半衰期比天然ADI的更长.重组ADI能抑制肿瘤细胞HepG2的生长,同时具有抑制HepG2细胞迁移的作用.A ADI （rADI） was cloned from Mycoplasma arginini by PCR and was then expressed in Escherichia coli. The rADI was purified with ion-exchange chromatogram to almost homogeneity. After refolding, the rADI was found to be similar in molecular weight （about 46 kD） and enzyme activity of converting L-arginine to citrulline （38 U/mg） as that of the the native. Based on the results from proteolysis assay, it was found that rADI exhibited more resistant to trypsin digestion compared with the wild-type. Eurther research indicated that this rADI had an inhibitory effect on the growth of HepG2 cells and the anti-angiogenic activity could be validated by the inhibition of migration of HepG2 cells.

关 键 词：精氨酸脱亚胺酶 抑制机制 肿瘤细胞生长 支原体 

分 类 号：Q786[生物学—分子生物学]