Mazzi Maurício V, Marcussi Silvana, Carlos Guilherme B, Stábeli Rodrigo G, Franco João J, Ticli Fábio K, Cintra Adélia C O, França Suzelei C, Soares Andreimar M, Sampaio Suely V
Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, FCFRP, USP, Ribeirao Preto-SP, Brazil.
Toxicon. 2004 Aug;44(2):215-23. doi: 10.1016/j.toxicon.2004.06.002.
A hemorrhagic metalloprotease, named BjussuMP-I, was isolated from Bothrops jararacussu snake venom by a combination of gel filtration on Sephacryl S-200 (0.01 M Tris-HCl, pH 7.6 buffer) and Phenyl Sepharose CL-4B chromatography (0.01 M Tris-HCl plus 4 M NaCl, pH 8.6 buffer, followed by a concentration gradient from 4 to 0 M NaCl at 25 degrees C in the same buffer). BjussuMP-I is a 60 kDa protein with a pI approximately 5.5, which induced hemorrhage after intradermal injection in mice, with a minimum hemorrhagic dose of 4.0 microg. The hemorrhagic activity of BjussuMP-I was totally abolished after incubation with a chelating agent (EDTA), corroborating the metal-dependency of this effect. BjussuMP-I shows proteolytic activity on casein and fibrinogen, although having an activity lower than that of crude B. jararacussu venom and the metalloprotease neuwiedase isolated from Bothrops neuwiedi snake venom. It was recognized by anti-neuwiedase antibodies, with a reaction of partial immunologic identity. BjussuMP-I also shows bactericidal activity against Escherichia coli and Staphylococcus aureus. This is the first report on the isolation and characterization of a high molecular weight hemorrhagic metalloprotease (BjussuMP-I) from B. jararacussu venom, which may play a relevant role in local and systemic bleeding which characterizes Bothrops envenomations.
一种名为BjussuMP-I的出血性金属蛋白酶，通过在Sephacryl S-200（0.01M Tris-HCl，pH 7.6缓冲液）上进行凝胶过滤和苯基琼脂糖CL-4B色谱法（0.01M Tris-HCl加4M NaCl，pH 8.6缓冲液，随后在25℃下在相同缓冲液中从4M到0M NaCl进行浓度梯度洗脱）相结合的方法，从巴西矛头蝮蛇毒中分离得到。BjussuMP-I是一种分子量为60 kDa、pI约为5.5的蛋白质，皮内注射到小鼠体内后会引起出血，最小出血剂量为4.0微克。BjussuMP-I与螯合剂（EDTA）孵育后，其出血活性完全丧失，这证实了这种作用对金属的依赖性。BjussuMP-I对酪蛋白和纤维蛋白原有蛋白水解活性，尽管其活性低于粗制的巴西矛头蝮蛇毒以及从纽氏矛头蝮蛇毒中分离得到的金属蛋白酶neuwiedase。它能被抗neuwiedase抗体识别，呈现部分免疫同一性反应。BjussuMP-I还对大肠杆菌和金黄色葡萄球菌具有杀菌活性。这是关于从巴西矛头蝮蛇毒中分离和鉴定一种高分子量出血性金属蛋白酶（BjussuMP-I）的首次报道，该蛋白酶可能在巴西矛头蝮蛇咬伤所致的局部和全身出血中发挥重要作用。
