广西产圆斑蝰蛇毒神经生长因子的分离纯化及鉴定

Isolation and Characterization of the Nerve Growth Factor from the Venom of Vipera russelli Siamensis

目的 从蝰蛇蛇毒中分离纯化神经生长因子 (NGF)。 方法 粗毒采用 CM5 2阳离子交换柱层析 ,DEAE Sephadex A - 5 0阴离子交换柱层析及 Sephadex G- 75凝胶过滤进行分离纯化。洗脱峰利用 PC1 2细胞测定活性 ,通过聚丙烯酰胺凝胶电泳 (PAGE) ,聚丙烯酰胺凝胶等电聚焦圆盘电泳 (EFG)等方法 ,进行理化性质的鉴定。 结果 从粗毒 6 .0 g中得到电泳纯的神经生长因子 3.6 4 m g,分子量为 36 k D,由两个亚基通过二硫键交联组成二体结构。等电点 p I为 6 .7,在 0 .5～ 5 .0μg/m L时 ,对 PC1 2细胞有良好的促进分化作用。

Objective To purifiy the nerve growth factor(NGF) from the venom of Vipera russelli siamensis. Methods The fraction with the activity of inducing PC12 cell differentiation was isolated and purified by CM52 ion exchange chromatography and DEAE Sephadex A 50 ion exchange chromatography and Sephadex G 75 gel filtration chromatography. Identification of NGF fraction by polyacrylamide gel electrophoresis(PAGE) and isoelectric focusing in PAGE(EFG). Results NGF 3 46 mg was purified from 6 0 g snake venom. The molecular weight was estimated to be 36 kD by SDS PAGE. The isoelectric point of NGF was pH 6 7. It consisted of two types polypeptide chains. In the dose of 0 5~5 0 μg/mL, the venom NGF can well induce PC12 cell differentiation. Conclusion NGF can be isolated and purified from the venom of Vipera russelli siamensis using the methods discussed.