神经诱向生长因子18kD蛋白的纯化

提取与纯化诱向生长因子是神经生物化学研究中的一个重要课题．采用自然系统凝胶电泳，分离周围神经损伤过程中产生的具有诱向生长作用的１８ｋＤ蛋白，再以等电聚焦凝胶电泳与神经组织联合培养，揭示了ｐＩ为５．２的１８ｋＤ蛋白具有诱神经生长的作用．经高效液相色谱仪分析获得较纯的１８ｋＤ诱向因子．蛋白／多肽测序仪检测，１８ｋＤ蛋白Ｎ端氨基酸序列为：ＰＥＰＡＷＳＡＰＡＰ．

Purification of 18 kD Protein of Neurochemotropic Factor

To detect novel tropic factor,further purification of active chemotropic factors is the key technique in the field of neurobiochemistry.The 18 kD protein,as a neurochemotropic factor,isolated from lesioned peripheral nerves by native polyacrylamide,was applied to investigate the possible active fraction of 18 kD protein and the strip of pI52 from isoelectrofocus gel.Then the electrophoresis of isoelectrofocus gel for the co culture with dorsal root ganglion showed that pI52 fraction of 18 kD protein possessed strong neurotropic activity.With the dilution of gradient conditions by HPLC,the pure 18 kD protein had been collected,and then the amino acid sequences of the 18 kD protein N terminal,PEPAWSAPAP had been analyzed on PVDF member after Western blotting by amino acid sequencer.The data presented here support the conclusion that the protocol is an effective way for the purification of active factor,and suggest that the pI52 fraction of 18 kD protein is responsible for the effect of chemotropism on nerve regeneration.