烙铁头(Trimeresurus mucrosquamatus)蛇毒的研究Ⅰ柱层析分离及酶活性与生物活性测定
Study on the Venom of Trimeresurus mucrosquamatus Ⅰ.Isolation of T.mucrosquamatus Venom and the Determination of its Biological and Enzymatic Activities
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摘要: 本文报导了用二乙氨基乙基葡聚糖凝胶A-50和氯化钠直线梯度洗脱方法，获得14个蛋白峰。测定了11种酶活性，含有精氨酸酯酶，蛋白水解酶，L-氨基酸氧化酶，5-核苷酸酶，腺三磷酸酶，磷酸单酯酶，磷酸二酯酶，核苷焦磷酸酶，磷脂酶A9种，不含核糖核酸酶，胆碱酯酶。对每个蛋白峰的生物活性测定表明，出血毒主要分布在12峰，其次为10—14峰。致死活性表现在1，12—14峰。1峰具有较强溶解纤维蛋白的活性。8，10—14峰具有明显的凝血酶样作用，酶活性的分布及其对凝血系统的作用接近一致。11—13峰能使血小板聚集。烙铁头毒蛇在我国分布很广，成都生物所两栖爬行动物研究室1974年作过报导。蛇毒的研究已被许多学者所重视，Suzuki在Anthony.T.Tu.编著的Venoms Chemistry and Moleculor Biology(1977)报导了该蛇毒含磷酸单酯酶，磷酸二酯酶，5-核苷酸酶及小分子活性多肽，1970年Ouyang报导含透明质酸酶，1963年Murata报导有蛋白水解酶活性。Ouyang等(1976—1978)分离纯化纤溶组分，并研究了该组分的理化性质。但对各组分的酶活性及生物活性测定尚未见报导。为充分利用这一丰富资源，我们对湖南产烙铁头蛇毒进行了柱层析分离和各组分的酶活性和生物活性测定，结果报导于下。Abstract: By means of DEAE-sephadex A-50 column chromatography,Trimeresurus mucrosquamqtus venom was separated into fourteen fractions.It contained proteolytic enzyme,5-nucleotidase,arginine esterase,etc.The enzymatic activities and biological activities of crude venom and each protein fraction were determined.The proteolytic enzyme was concentrated in the fractions 1,2,4,7,and 8.Among them the fraction 1 had the highest activity of proteolytic enzyme.Fraction 2 possessed phospholipase A activity,but not the others.Fraction 7-14 possessed arginine esterase activity.Fractions 10-14 possessed hemorrhagic principles and among them fraction 12 was the highest.Fractions 1,12-14 possessed lethal toxicity.Fractions 11-13 showed the effect of platelet aggregation.
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