蕲蛇蛇毒抗凝血因子的分离纯化与特性

Purification and Characterization of An Anticoagulant Factor from the Venom of Agkistuodon acutus

目的 寻找蕲蛇蛇毒中的抗凝血因子。 方法 利用硫酸铵沉降、阴离子交换层析、阳离子交换层析及高效液相色谱层析 ,从蕲蛇蛇毒中分离纯化到一个抗凝血因子。 结果 纯化的这一组份在PAGE、SDS-PAGE上均呈单一区带 ,分子量约为 2 5 .4k D,由两条分子量分别为 1 5 .0 k D和 1 6.0 k D的肽链通过二硫键连接在一起。这一组份在体外显著地延长血浆复钙时间和凝血酶原时间 ,但不延长牛凝血酶时间 ,也不具有磷脂酶 A2 活性、纤溶活性和出血活性。

Objective To investigate the anticoagulant factor from the venom o Agkistuodon acutus Methods Using ammonia sulfate sedimentation,anion exchange chromatography (DEAE Sephadex A 50),followed by cation exchange chromatography (CM Sephadex C 50) and anion exchange chromatography (POROS 20 PI) to purifying the anticoagulant factor from the venom of Agkistuodon acutus Results The fraction was homogeneous detemined by SDS polyacrylamid gel electrophoresis and polyacrylamide gel electrophoresis,which was of apparent molecular weight of about 25 4kD under nonreduced conditions,and could be reduced to about 15 0kD and 16 0kD residues respectively by mercaptoethanol In vitro,it remarkably prolonged the plasma recalcification time and the plasma prothrombin time(PPT),but did not dissolve fibrin and could not inhibit bovine thrombin Furthermore,it did not show both the phospholipase A 2 activity and the hemorrhagic antivity Conclusions There were a new anticoagulant factor in the venom of Agkistuodon acutus