Abstract
The bioactivities and proteinous compositions of venoms collected by two different methods from Latrodectus tredecimguttatus were analyzed and compared. Most of proteinous venom components were high-molecular-mass acidic proteins (>10<sup>4</sup>). Compared with the venom obtained from dissected venom gland (DGV), the venom obtained by electrical stimulation (ESV) contained more high molecular mass proteins, but the venom proteins and peptides with low molecular mass (<10<sup>4</sup>) in the preparations were very similar. Intraperitoneal injection of the two venoms in mice gave rise to similar poisoning symptoms and the LD<sub>50</sub> values of ESV and DGV were (0.16±0.03) mg/kg and (0.39±0.05) mg/kg, respectively. LD<sub>50</sub> values of ESV and DGV in cockroaches (Periplaneta Americana) were 1.87 μg/g and 2.32 μg/g, respectively. ESV was able to block neuromuscular transmission in the mouse phrenic nerve-hemidiaphragm preparation within (25.0±2.2) min at a concentration of 3.2×10<sup>-6</sup>g/mL, whereas DGV blocked neuromuscular transmission within (45.7±1.8 )min at the same concentration. The fraction of the venom containing proteins and peptides with molecular mass <10<sup>4</sup> did not obviously affect the transmission. Those results demonstrated that Latrodectus tredecimguttatus venom was a mixture rich in larger proteins rather than smaller proteins and peptides; the mammalian toxicities of the venom were due primarily to the high molecular weight acidic proteins rather than low molecular weight peptides; the active components contained in ESV and DGV were similar but the content in ESV was higher than that in DGV.
Highlights
The bioactivities and proteinous compositions of venoms collected by two different methods from Latrodectus tredecimguttatus were analyzed and compared
Compared with the venom obtained from dissected venom gland (DGV), the venom obtained by electrical stimulation (ESV) contained more high molecular mass proteins, but the venom proteins and peptides with low molecular mass (
The fraction of the venom containing proteins and peptides with molecular mass
Summary
Venom Properties of the Spider Latrodectus tredecimguttatus and Comparison of Two Venom-Collecting Methods Abstract: The bioactivities and proteinous compositions of venoms collected by two different methods from Latrodectus tredecimguttatus were analyzed and compared. 间斑寇蛛(Latrodectus tredecimguttatus)(又名 黑寡妇蜘蛛)毒液是由多种活性成分组成的混合 物,是间斑寇蛛捕获猎物和防御敌害的有效武器。 在过去的几十年中,间斑寇蛛因频繁引发伤害人、 畜的事件及其毒液中大量具有药用前景的生物活 性成分的存在而引起了人们的广泛关注(Ushkaryov et al, 2004; Isbister & White, 2004)。研究间斑寇蛛毒 液的组成和性质对于阐明蛛毒中毒的机理、防治蜘 蛛伤害及活性毒素成分的开发利用均具有重要的 意义。将毒腺匀浆应用于龙虾神经-肌肉标本时,发 现它能先增强然后减弱兴奋性和抑制性突触后电 位,表明粗毒液中含有作用于龙虾神经肌肉接头突 触前膜的成分(Kawai et al, 1972; Burmistrov et al, 1997)。将毒腺匀浆应用于青蛙神经-肌肉标本时, 发现它能够导致小终板电位频率的增加及神经末 端突触囊泡的减少(Clark et al, 1972; Fesce et al, 1986)。迄今为止,已从间斑寇蛛毒液中分离鉴定到 包括Latrotoxins在内的 7 种主要的毒性成分。所有 研究过的Latrotoxins类毒素是一类大分子酸性蛋白 质,相对分子质量范围分布在 1.1×105~1.4×105之 间,等电点分布在 5.5~6.0 之间;能导致神经末端 递质分泌的显著增加,且对脊椎动物、昆虫或甲壳 类动物有着不同的专一性(Ushkaryov et al, 2004)。 其中被研究得最多的是α-latrotoxin, 在有或无Ca2+ 的条件下都可以通过诱导突触递质的大量释放而 对脊椎动物产生致命的影响(Clark et al, 1972; Fesce et al, 1986; Ceccarelli & Hurlbut, 1980; Cavalieri et al, 1990; Orlova et al, 2000; Volynski et al, 2003; Lajus et al, 2006) 。 此 外 , Akhunov et al (1992) 和 Akhunov(1996)对间斑寇蛛(L.tredecimguttatus)毒 液中的激酶和两种具有血管舒缓激肽增效作用的 多肽进行了研究。他们确定了这种激酶是一种巯基 内肽酶,这种酶能够催化切断肽内的脯氨酸羧基端 的肽键。而对大鼠的实验结果表明,两种血管舒缓 激肽增效多肽能够延长血管舒缓激肽的抑制作用、 刺激细胞释放组氨酸和降低血压。Volkova et al (1995)从间斑寇蛛粗毒液中分离出了两种低相对分 子质量蛋白质LMWP和LMWP2,实验证明这两种 物质对脊椎动物和蜚蠊(Periplaneta Americana)均 无毒性。
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