摘要
本文用SP-SephadexC-25柱层析法分离广西眼镜王蛇蛇毒,得到19个组分,其中7个组分有毒性。对毒性较强,含量较高的4个毒性组分(即神经毒素Ⅴ、Ⅶ、Ⅷ和Ⅹ)用SephadexG-50凝胶过滤法进行纯化,并对神蛇经毒素Ⅶ和Ⅹ用CM-SephadexC-50柱层析法做了进一步纯化。经聚丙烯酰胺凝胶电泳(PAGE)鉴定,这4个神经毒素均为单一条带。用SDS-PAGE法测得神经毒素Ⅴ、Ⅶ、Ⅷ和Ⅹ的分子量分别为7800,6700,8400和8000,并用它们进行了小鸡颈二腹肌神经—肌肉接头传递的阻遏试验,结果表明这4个神经毒素均有阻遏神经—肌肉接头传递的作用,而且阻遇作用基本不可逆,属于突触后神经毒素。
From the venom of King Cobra, nineteen fractions were isolated by column chromatography on SP-SephadexC-25. Of the nineteen fractions, seven were toxic. Four toxic fractions (i. e. neurotoxin Ⅴ, Ⅵ, Ⅷ and Ⅹ)with stronger toxicities and higher contents were purified by gel filtration SephadexG-50,and neurotoxin Ⅶ and Ⅹ were further purified by column chromatography on CM-SephadexC-50. All the four neurotoxins showed a single band on polyacrylamide gel electrophoresis(PAGE). The molecular weight of neurotoxin Ⅴ,Ⅶ,Ⅷ and Ⅹ was estimated to be 7 800, 6 700, 8 400 and 8 000 respectively by SDS-PAGE, and the experiments of neuromuscular junction transmission in chick biventer cervicix were made with the four neurotoxins. The results showed that all the four had the effect in blocking the transmission of neuromuscular junction.Furthermore, the blocking effect was essentially irreversible. They belonged to the postsynapse neurotoxin.
关键词
眼镜王蛇
蛇毒
神经毒素
柱层析
King Cobra venom
neurotoxins
column chromatography
blocking experiment
