虎纹捕鸟蛛粗毒双向凝胶电泳分析及部分蛋白质点的N端序列测定

Two-dimensional Gel Electrophoresis of the Crude Venom from the Spider Selenocosmia huwena and the Identification of Protein Spots by Microsequencing

采用固相 p H梯度等电点聚焦 - SDS双向聚丙烯酰胺凝胶电泳对虎纹捕鸟蛛粗毒进行了分析 ,通过考马斯亮蓝与银染法显色 ,电脑软件识别出约 30 0个蛋白质点 .约有 35个含量较高的蛋白质点分布在分子量 1 0 k D以下区域 ,通过印迹法将凝胶上蛋白质点转移到 PVDF膜上以后 ,对上述分子量 1 0 k D以下的组分进行了 N端序列测定 ,鉴定到了虎纹捕鸟蛛毒素 HWTX- ,HWTX- ,HWTX- 和 SHL- 1在凝胶上的位置 ,同时发现了 5种新的肽类毒素组分 .

Crude venom of the spider Selenocosmia huwena was separated by two dimensional gel electrophoresis(2D PAGE),with the separation in the first dimension on a wide range immobilized pH(3-10)gradient.Over 300 protein spots were presented on a silver stained 2D gel.About 35 spots with relatively high concentration were located on the region below molecular weight of 10 kD.The protein spots were electro transferred to polyvinylidene difluoride(PVDF)membrane and the low molecular weight spots were analyzed by N terminal protein microsequencing.Four known toxic peptides:HWTX Ⅰ,HWTX Ⅱ,HWTX Ⅳ and SHL Ⅰ were identified and five of new components were found among these protein spots.