江浙蝮蛇(Agkistrodon halys Pallas)蛇毒神经生长因子的分离纯化和特性

神经生长因子（ｎｅｒｖｅ ｇｒｏｗ ｔｈ ｆａｃｔｏｒ， Ｎ Ｇ Ｆ）是第一个被发现，也是迄今为止研究得最为清楚的一种神经营养因子 利用 Ｐ Ｃ１２ 细胞生物活力测定为跟踪检测手段，分别经过 Ｃ Ｍ Ｓｅｐｈａｒｏｓｅ Ｃ Ｌ ６ Ｂ、 Ｓｅｐｈａｄｅｘ Ｇ ７５ 及 Ｆ Ｐ Ｌ Ｃ ｍ ｏｎｏ Ｓ层析，从３０ ｇ 江浙蝮蛇粗毒中分离纯化到２００ μｇ Ｎ Ｇ Ｆ，纯化倍数高达１０５经 Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ 测定，该蛋白分子量为 ２６ ｋ Ｄ，由两个亚基通过二硫键交联组成二体形式等电聚焦显示其等电点为６７，与氨基酸组成分析结果相吻合 江浙蝮蛇神经生长因子的生物活力水平与小鼠２５ Ｓ Ｎ Ｇ Ｆ相当，在１～１００ μｇ／ Ｌ 的浓度范围内维持 Ｐ Ｃ１２ 细胞在无血清条件下的存活

Isolation and Characterization of the Nerve Growth Factor from the Crude Venom of Agkistrodon halys Pallas

Nerve growth factor(NGF)is the first and the best characterized neurotrophic factor Since NGF is required for normal survival of adult sympathetic and sensory neurons,it has been investigated for use as a therapeutic agent in neurologic diseases,especially Alzheimer′s disease(AD) Snake venom is one of the major source of NGF,however,NGFs of venoms from different families of snakes show differences in properties NGF from the venom of Agkistrodon halys Pallas was sequentially purified by CM Sepharose CL 6B ion exchange chromatography,Sephadex G 75 gel filtration and mono S column chromatography on FPLC system The NGF in the venom of A halys Pallas is so scare that only 200 μg NGF was obtained from 30 g crude venom On SDS PAGE,the purified NGF migrated as 26 kD dimer,with subunits convalently linked by a disulphide bond Analytical isoelectric focusing revealed the protein with p I about 6 7 Its biological activity was comparable to that of mouse 2 5 S NGF The NGF preparation of A halys Pallas venom at the concentration interval 1-100 μg/L can rescue cultured PC12 cells from apoptosis