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蝮蛇突触前毒素的结晶及其物化性质的鉴定

THE CRYSTALLIZATION AND PHYSICOCHEMICAL CHARACTERIZATION OF AGKISTRODOTOXIN

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【作者】 张景康江明恃吴健永徐科陈远聪武祥福

【Author】 ZHANG JING-KAN JIANG MING-SHI SHU KE (Shanghai Institute of Physiology, Academia Sinica) CHEN YUAN-CHUNG WU XIAN-FU (Shanghai Institute of Biochemistry, Academia Sinica)

【机构】 中国科学院上海生理研究所北京大学中国科学院上海生物化学研究所中国科学院上海生物化学研究所 79届毕业生

【摘要】 江苏和浙江省一带所产的蝮蛇(Agkistrodon halys Pallas)毒,按前文所描述方法,分离出二个主要神经毒组分。经生理学鉴定其中一个具有选择性地阻断神经肌肉接头突触前传递过程的作用。用等电聚焦板电泳从该组分制备出经聚丙烯酰胺圆盘电泳鉴定为一均一条纹的突触前毒素,定名为蝮蛇神经毒素(agkistrodotoxin),蝮蛇神经毒素对小白鼠(腹腔注射)的最小致死量为每公斤体重55微克,在90℃下保温20分钟毒性不变,用中性连续或碱性不连续SDS圆盘电泳测定的分子量为13,400,不含亚基,用等电聚焦圆盘电泳测得等电点为6.9,在12℃对34%饱和硫酸铵(pH7.0)透析可得长方形晶体。关于蝮蛇神经毒素的其他生理特性,以及蝮蛇毒中其他神经毒素的纯化和鉴定将在另文发表。

【Abstract】 The crude venom of the pit viper (Agkistrodon halys Pallsr) collected in Jiangsu and Zhejiang provinces was analysed at first by anion exchange column chromatography on DEAE-cellulose. The main neurotoxic component obtained then underwent a further analysis by gel filtration on Sephadex G-50 and a component with prosynaptio neurotoxicity could be isolated, which was treated as a crude agkistrodotoxin in some of our physiological and biochemical experiments. The further purification of the crude agkistrodotoxin was carried out by preparative isoelectric focusing on polyacrylamide gel places. It was shown that the purified agkistrodotoxin (ATX) migrated as a single band in disk polyacrylamide electrophoresis.The minimal lethal dose (i. p.) of ATX was about 55r per kg body weight in the mouse. There was no change in neurotoxioity of ATX after subjection to 90℃ for 20 min, so it was rather heat resistant. No subunit was observed in the molecule of ATX, the molecular weight was found to be 13400, as determined by electrophoresis on SDS-polyacrylamide gel. The isoelectric point of ATX was pH6.9. The crystals of ATX were obtained by dialysing against 34% saturated ammonium sulfate solution at pH 7.0 and 12℃.

  • 【文献出处】 Acta Biochimica et Biophysica Sinica ,生物化学与生物物理学报 , 编辑部邮箱 ,1981年03期
  • 【被引频次】5
  • 【下载频次】56
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