Cecchini Alessandra L, Marcussi Silvana, Silveira Lucas B, Borja-Oliveira Caroline R, Rodrigues-Simioni Léa, Amara Susan, Stábeli Rodrigo G, Giglio José R, Arantes Eliane C, Soares Andreimar M
Departamento de Física e Química, FCFRP, USP, Ribeirão Preto-SP, Brazil.
Comp Biochem Physiol A Mol Integr Physiol. 2005 Jan;140(1):125-34. doi: 10.1016/j.cbpb.2004.11.012.
The venoms of Micrurus lemniscatus carvalhoi, Micrurus frontalis frontalis, Micrurus surinamensis surinamensis and Micrurus nigrocinctus nigrocinctus were assayed for biological activities. Although showing similar liposome disrupting and myotoxic activities, M. frontalis frontalis and M. nigrocinctus nigrocinctus displayed higher anticoagulant and phospholipase A2 (PLA2) activities. The latter induced a higher edema response within 30 min. Both venoms were the most toxic as well. In the isolated chick biventer cervicis preparation, M. lemniscatus carvalhoi venom blocked the indirectly elicited twitch-tension response (85+/-0.6% inhibition after a 15 min incubation at 5 microg of venom/mL) and the response to acetylcholine (ACh; 55 or 110 microM), without affecting the response to KCl (13.4 mM). In mouse phrenic nerve-diaphragm preparation, the venom (5 microg/mL) produced a complete inhibition of the indirectly elicited contractile response after 50 min incubation and did not affect the contractions elicited by direct stimulation. M. lemniscatus carvalhoi inhibited 3H-L-glutamate uptake in brain synaptosomes in a Ca2+-, but not time, dependent manner. The replacement of Ca2+ by Sr2+ and ethylene glycol-bis(beta-aminoethyl ether) (EGTA), or alkylation of the venom with p-bromophenacyl bromide (BPB), inhibited 3H-L-glutamate uptake. M. lemniscatus carvalhoi venom cross-reacted with postsynaptic alpha-neurotoxins short-chain (antineurotoxin-II) and long-chain (antibungarotoxin) antibodies. It also cross-reacted with antimyotoxic PLA2 antibodies from M. nigrocinctus nigrocinctus (antinigroxin). Our results point to the need of catalytic activity for these venoms to exert their neurotoxic activity efficiently and to their components as attractive tools for the study of molecular targets on cell membranes.
对米氏珊瑚蛇(Micrurus lemniscatus carvalhoi)、额部珊瑚蛇(Micrurus frontalis frontalis)、苏里南珊瑚蛇(Micrurus surinamensis surinamensis)和黑环珊瑚蛇(Micrurus nigrocinctus nigrocinctus)的毒液进行了生物活性测定。尽管这几种毒液表现出相似的脂质体破坏和肌毒性活性,但额部珊瑚蛇和黑环珊瑚蛇的抗凝和磷脂酶A2(PLA2)活性更高。后者在30分钟内引起更高的水肿反应。这两种毒液也是毒性最强的。在离体鸡双腹肌制备中,米氏珊瑚蛇毒液阻断了间接引发的抽搐-张力反应(在5微克毒液/毫升中孵育15分钟后抑制率为85±0.6%)以及对乙酰胆碱(ACh;55或110微摩尔)的反应,而不影响对氯化钾(13.4毫摩尔)的反应。在小鼠膈神经-膈肌制备中,毒液(5微克/毫升)在孵育50分钟后完全抑制了间接引发的收缩反应,且不影响直接刺激引发的收缩。米氏珊瑚蛇毒液以依赖钙离子但不依赖时间的方式抑制脑突触体对3H-L-谷氨酸的摄取。用锶离子和乙二醇双(β-氨基乙醚)(EGTA)替代钙离子,或用对溴苯甲酰溴(BPB)对毒液进行烷基化处理,均可抑制3H-L-谷氨酸的摄取。米氏珊瑚蛇毒液与突触后α-神经毒素短链(抗神经毒素-II)和长链(抗银环蛇毒素)抗体发生交叉反应。它还与来自黑环珊瑚蛇的抗肌毒性PLA2抗体(抗黑环蛇毒素)发生交叉反应。我们的结果表明,这些毒液需要催化活性才能有效地发挥其神经毒性活性,并且其成分是研究细胞膜上分子靶点的有吸引力的工具。