Curin-Serbec V, Délot E, Faure G, Saliou B, Gubensek F, Bon C, Choumet V
Unité des Venins, Institut Pasteur, Paris, France.
Toxicon. 1994 Nov;32(11):1337-48. doi: 10.1016/0041-0101(94)90406-5.
Crotoxin and ammodytoxin A are snake venom neurotoxic phospholipases A2. Polyclonal antibodies against three synthetic peptides selected from the C-terminal part of the primary structure of ammodytoxin A were tested by ELISA for their interaction with crotoxin and its subunits, CA and CB. All three antipeptide antibodies reacted specifically with corresponding parts of ammodytoxin A and CB, either native or reduced. Conversely, polyclonal antibodies produced against ammodytoxin A and CB reacted strongly with all three peptides, suggesting that they constitute at least a part of natural epitopes in both proteins. All antipeptide antibodies reacted also with the corresponding peptides derived from CB by cyanogen bromide cleavage. The biological activity of the immune complexes was tested. No significant change in the enzymatic activity of CB, ammodytoxin A or crotoxin was observed with any of the three antipeptide antibodies. These antibodies were, however, able to protect mice against the lethal potency of CB and to prolong survival time of mice injected with crotoxin. These antipeptide antibodies were assayed in vitro for their protective effect against the action of CB or crotoxin on synaptosomes from Torpedo marmorata electric organ. They partly inhibited the acetylcholine release induced by both proteins. These results indicate that the C-terminal part of CB is likely to be involved in the pharmacological action of crotoxin.
响尾蛇毒素和锯鳞蝰毒素A是蛇毒神经毒性磷脂酶A2。通过酶联免疫吸附测定(ELISA)测试了针对从锯鳞蝰毒素A一级结构的C末端部分选择的三种合成肽的多克隆抗体与响尾蛇毒素及其亚基CA和CB的相互作用。所有三种抗肽抗体都与天然或还原形式的锯鳞蝰毒素A和CB的相应部分发生特异性反应。相反,针对锯鳞蝰毒素A和CB产生的多克隆抗体与所有三种肽都发生强烈反应,表明它们构成了这两种蛋白质中天然表位的至少一部分。所有抗肽抗体也与通过溴化氰裂解从CB衍生的相应肽发生反应。测试了免疫复合物的生物学活性。用三种抗肽抗体中的任何一种都未观察到CB、锯鳞蝰毒素A或响尾蛇毒素的酶活性有显著变化。然而,这些抗体能够保护小鼠免受CB的致死效力,并延长注射响尾蛇毒素的小鼠的存活时间。在体外测定了这些抗肽抗体对CB或响尾蛇毒素对电鳐电器官突触体作用的保护作用。它们部分抑制了这两种蛋白质诱导的乙酰胆碱释放。这些结果表明,CB 的C末端部分可能参与响尾蛇毒素的药理作用。