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目的 被尖吻蝮蛇 （ Ｄｉｅｎａｇｋｉｓｔｒｏｄｏｎ ａｃｕｔｕｓ） 咬伤会引起严重的出血， 对蛇毒出血毒素的研究有利于治疗蛇伤出血药物筛选。 方法 采用 Ｓｅｐｈａｄｅｘ Ｇ７５， Ｄ Ｅ Ａ Ｅ Ｓｅｐｈａｄｅｘ Ａ５０， Ｓｅｐｈａｄｅｘ Ｇ２００ 和两次 Ｐ Ｂ Ｅ 聚焦层析纯化。 Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ 电泳和等电聚焦电泳测定纯化样品的纯度和等电点。氨基酸组成用自动氨基酸分析仪测定。以小鼠背部皮下注射部位出血斑的面积来确定最小出血剂量和常规的方法测定酶活性。结果 从尖吻蝮蛇毒中纯化到一个相对分子量为５６ ０００ 的出血毒素 （ Ｄａ Ｈ Ｔ３）， 经氨基酸组成测定计算，它由 ４８７ 个氨基酸残基组成。此成分在 Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ上显示出一条均一的蛋白染色带， 其ｐ Ｉ为５５０。该出血成分的最小出血剂量是 ２６μｇ， 具有蛋白水解酶活力， 其活力为 ３６８， 但没有精氨酯酶和磷脂酶 Ａ２ 活力。当加入 Ｅ Ｄ Ｔ Ａ 螯合剂去除金属离子后， 它们的出血活力和蛋白水解酶活力均丧失。 结论 这是从大陆尖吻蝮蛇毒中获得的一个新的出血金属蛋白酶 （ Ｄａ Ｈ Ｔ３）。
Objective Bite by Dienagkistrodon acutus can cause serious bleeding, and the study of venom toxins from snake venom is helpful for the treatment of snakebite hemorrhage. Methods Sephadex G75, D E A ESephadex A50, Sephadex G200 and twice P B E focused chromatography were used for purification. S D S P A G E electrophoresis and isoelectric focusing electrophoresis to determine the purity and isoelectric point of the purified sample. Amino acid composition with automatic amino acid analyzer. The area of bleeding spots at the subcutaneous injection site on the back of the mice was used to determine the minimum bleeding dose and the conventional method was used to determine the enzyme activity. Results Purified from Agkistrodon acutus venom to a relative molecular weight of 56 000 hemorrhagic toxin (Da H T 3), calculated by the amino acid composition calculation, which consists of 487 amino acid residues. This component showed a uniform protein band on SDS-PAGE, with a p I of 550. The hemorrhagic component of the minimum bleeding dose is 2 6μg, with proteolytic activity, the activity of 3 68, but no arginase and phospholipase A2 activity. When E D T A chelating agent was added to remove metal ions, their hemorrhagic activity and proteolytic activity were lost. Conclusion This is a new hemorrhagic metallo-protease (Da H T-3) obtained from the venomous Agkistrodon acutus.