Isolation And Activity Analysis Of The Toxic Components From Latrodectus Tredecimguttatus Spiderlings

Latrodectus tredecimguttatus, "black widow spider" or "black venomous spider", is one of medium-sized venomus spiders, which belong to Phylum Arthopoda, Arachnidea, Araneae, Theridiidae, genus Latrodectus. L. tredecimguttatus spider is one of the most poisonous spiders known in the world. In the past decades, the related scientific reseachers at home and abroad have made detailed studies on the venom secreted by the venom glands of adult spiders. However, in contrat to that the snake venom is found in only in the venom glands, the venom of L. tredecimguttatus is distributed in, besides the venom glands, the other parts of the body including legs and abdomen. Up to now, there have not been any reports on the systematic study on these toxins in the spider body. In the present paper, the toxic component in the newborn spider body were extracted, purified and characterized systematically. The soluble protein content of the lyophilized extract mearured in Bradford method was 27.72%. Electrophysiological experiments showed that the extract could block the neuromusclar transmission in isolated mouse phrenic nerve-hemidiaphragm prapration. When the concentration of the added extract was 80μg/ml, the transmission was completely blocked within about 16.0 min. Gel filtration chramotography of the extract resulted in 6 major fractions and SDS-PAGE demonstrated that the high-molecular-weight proteins were well separated from the low-molecular-weight components. The further electrophysiological analsyses using mouse phrenic nerve-hemidiaphragm prapration indicated that the fractions of peak 1 and peak 2 containing proteins with molecular weight greater than 10 kDa could block the neuromusclar transmission. The components in fraction of peak 1 completely blocked the neuromusclar transmission within 16.0±1.5 min in the concentration of 2.03 mg/ml, and this effect could not be eliminated by washing with Tyrode's solution. The components in fraction of peak 2 completely blocked the neuromusclar transmission within 15.0±2.0 min in the concentration of 2.99 mg/ml, and this effect could not be eliminated by washing with Tyrode's solution. However the components with molecular weight smaller than 10 kDa could not block the neuro musclar transmission completely, and even increasing the concentration to 5mg/ml, it could only block partial transmission, suggesting that the active components toxic to nerve system of mammals present primarily in the fractions containing proteins with molecular weight greater than 10 kDa.