Ami Amina, Oussedik-Oumehdi Habiba, Laraba-Djebari Fatima
USTHB, Faculty of Biological Sciences, Laboratory of Cellular and Molecular Biology, Bab Ezzouar, Algiers, Algeria.
J Biochem Mol Toxicol. 2017 Feb;31(2). doi: 10.1002/jbt.21835. Epub 2016 Sep 2.
A dermonecrotic metalloproteinase (CcD-II) was isolated from C. cerastes venom. Venom fractionation was performed using three chromatographic steps (molecular exclusion on Sephadex G-75, ion-exchange on DEAE-Sephadex A-50, and reversed-phase high-performance liquid chromatography on C8 column). CcD-II presented an apparent molecular mass of 39.9 kDa and displayed a dermonecrotic activity with a minimal necrotic dose of 0.2 mg/kg body weight. CcD-II showed proteolytic ability on casein chains and on α and β fibrinogen chains that was inhibited by ethylenediamine tetraacetic acid and 1,10-phenanthroline while remained unaffected by phenylmethylsulphonyl fluoride and heparin. CcD-II displayed gelatinase activity and degraded extracellular matrix compounds (type-IV collagen and laminin). These results correlated with histopathological analysis showing a complete disorganization of collagenous skin fibers. These data suggested that CcD-II belongs to P-II class of snake venom metalloproteinase. The characterization of venom compounds involved in tissue damage may contribute in the development of new therapeutic strategies in envenomation.
从锯鳞蝰毒液中分离出一种皮肤坏死金属蛋白酶（CcD-II）。毒液分级分离通过三个色谱步骤进行（在Sephadex G-75上进行分子排阻、在DEAE-Sephadex A-50上进行离子交换以及在C8柱上进行反相高效液相色谱）。CcD-II的表观分子量为39.9 kDa，表现出皮肤坏死活性，最小坏死剂量为0.2 mg/kg体重。CcD-II对酪蛋白链以及α和β纤维蛋白原链具有蛋白水解能力，这种能力受到乙二胺四乙酸和1,10-菲啰啉的抑制，而不受苯甲基磺酰氟和肝素的影响。CcD-II具有明胶酶活性，并能降解细胞外基质化合物（IV型胶原和层粘连蛋白）。这些结果与组织病理学分析相关，该分析显示皮肤胶原纤维完全紊乱。这些数据表明CcD-II属于蛇毒金属蛋白酶的P-II类。对参与组织损伤的毒液化合物进行表征可能有助于开发新的蛇伤治疗策略。
