浙江尖吻蝮蛇蛇毒类凝血酶纯化及促凝活性

Purification and clotting activity of a thrombin-like enzyme from Zhejiang agkistrodon acutus venom

摘要: 采用柱色谱等纯化方法从浙江尖吻蝮蛇凝血酶中提取获得一种类凝血酶（TQ-1），采用RP-HPLC、MALDI-TOF、SDS-PAGE、等电聚焦电泳、Edman降解法对其进行了结构确证，并对其体内、外凝血活性进行测定。纯化所得TQ-1纯度98.5%，相对分子质量为30 522.95,SDS-PAGE分析其为一种单链蛋白，等电点为4.5，N末端氨基酸序列为VIGGNECDTNEHRFL。体外凝血活性为900U/mg，小鼠体内凝血试验结果表明：TQ-1凝血活性与阳性对照药巴曲亭相似。TQ-1为一种未见报道的尖吻蝮蛇类凝血酶，具有较高的开发价值。

Abstract: A thrombin-like enzyme named TQ-1 was purified by column chromatography from Zhejiang agkistrodon acutus venom.The structure of TQ-1 was studied by RP-HPLC,Q-TOF,SDS-PAGE,Edman degradation and isoelectric focus;clotting activity of TQ-1 was studied in vivo and vitro.The purity of TQ-1 by was 98.5%,the molecular weight was 30 522.95,the isoelectric point was 4.5,the N-terminal amino acids sequence was VIGGNECDTNEHRFL,the specific activity in vitro was 900U/mg,and the clotting activity in vivo was similar to that of the positive control.TQ-1 is a new type of thrombin-like enzyme with a high medicinal value.