Fuly A L, Francischetti I M, Zingali R B, Carlini C R
Departamento de Bioquímica, Universidade Federal do Rio de Janeiro, Brasil.
Braz J Med Biol Res. 1993 May;26(5):459-63.
Screening of the biochemical-pharmacological properties of the crude venom from the snake Lachesis muta indicated the presence of phospholipase A2 (PLA2; 5260 U/mg protein), procoagulant (2630 U/mg protein), platelet aggregating (43 U/mg protein) and caseinolytic activities (6670 U/mg protein). These activities were separated by filtration of the crude venom on Sephacryl S-200. The material containing PLA2 activity was further fractioned by DEAE-cellulose ion exchange chromatography into four active fractions (F-I to F-IV, containing 1.7, 1.2, 0.3, and 0.05% of the crude venom protein, respectively) by stepwise elution with buffers of increasing ionic strength. All fractions presented a molecular weight of approximately 15,000 and isoelectric points in the range pH 4.6-6.0. In addition to their indirect hemolytic activity, the partially purified fractions inhibited platelet aggregation induced either by collagen or thrombin. p-Bromophenacyl bromide-treated fractions lost both phospholipase A2 activity and their inhibitory effect on collagen-induced platelet aggregation.
对矛头蝮蛇粗毒的生化药理特性进行筛选,结果表明其含有磷脂酶A2(PLA2;5260 U/mg蛋白质)、促凝剂(2630 U/mg蛋白质)、血小板聚集活性(43 U/mg蛋白质)和酪蛋白溶解活性(6670 U/mg蛋白质)。通过在Sephacryl S - 200上对粗毒进行过滤,可分离出这些活性成分。含有PLA2活性的物质通过DEAE - 纤维素离子交换色谱进一步分级,采用离子强度递增的缓冲液分步洗脱,得到四个活性组分(F - I至F - IV,分别含有粗毒蛋白质的1.7%、1.2%、0.3%和0.05%)。所有组分的分子量约为15,000,等电点在pH 4.6 - 6.0范围内。除了具有间接溶血活性外,部分纯化的组分还能抑制由胶原或凝血酶诱导的血小板聚集。经对溴苯甲酰溴处理的组分既失去了磷脂酶A2活性,也失去了对胶原诱导的血小板聚集的抑制作用。