摘要
目的 :从猴的肾上皮细胞 (Vero- E6 )内提取具有促进细胞生长作用的 3-磷酸甘油醛脱氢酶 (G3PD)的修饰蛋白 (m odifier protein,MP)。方法 :将培养液中 K+的浓度降低到 3.2 mmol/ L,在 Vero- E6细胞内将产生 G3PD的修饰蛋白 ,用亲和层析及阴离子高效液相层析纯化而获取这种修饰蛋白。结果 :蛋白的分子量大约为 6 2 k Da,Westem及免疫吸附实验证实 :随着细胞外液中 K+的浓度降低 ,该种蛋白也相应增多 ,在低 k溶液 2 h后达最大值 ,与 G3PD的活性相平行。结论 :从猴的肾上皮 Vero- E6细胞分离出 G3PD的修饰蛋白 ,有促进 G3PD的活性。
Objective: Our aim was to purity the modifier protein of glyceraldehyde 3 phosphate dehydrogenase (G3PD) from African green monkey Vero E6 line. Methods:Exposure of Vero E6 cells to medium with a reduced K concentration (3.2 mmol/L) stimulated the growth and activation of G3PD. The increase of enzyme activity was mediated by a cytosolic modifier protein that was purified using affinity and anion exchange high performance liquid chromatograph. Results:The apparent molecular mass of the protein was 62 kDa. Western blotting and quantiative enzyme linked immunosorbent assay showed that the amount of modifier protein increased progressively for 2 hours in cells exposed to low K + medium, and then returned to the control value, a kinetic profile similar to that the modifier protein is a constituent of renal epithelial cells and accummulated transiently in the low K + mitogenic signal. Conclusion: We obtained a modifer protein from monkey kidney epithelial cells (Vero E6). It could activate G3PD and cell growth.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2001年第1期31-32,40,共3页
Journal of China Medical University
基金
卫生部科研基金资助项目 !96-1-2 11