Laboratory of Toxicology, University of Leuven, Campus Gasthuisberg, Leuven, Belgium.
Proteomics. 2012 Jan;12(2):313-28. doi: 10.1002/pmic.201100409. Epub 2012 Jan 4.
Venom from the scorpion Pandinus cavimanus was obtained by electrical stimulation of the telson (stinger). Total venom was toxic to crickets at 7-30 μg and a paralysis or lethal effect was observed at 30 μg of venom (death at 1.5 μg/mg of cricket). Electrophysiological analyses showed cytolytic activity of total venom on oocytes at 7 μg. HPLC allowed separation of the venom components. A total of 38 fractions from total venom were tested on voltage-gated Na(+) and K(+) channels. Some fractions block K(+) currents in different degrees. By using MS analysis, we obtained more than 700 different molecular masses from telson and venom fractions (by LC-MS/MS and MALDI-TOF MS analyses). The number of disulfide bridges of the telson components was determined. A cDNA library from P. cavimanus scorpion was constructed and a random sequencing screening of transcripts was conducted. Different clones were obtained and were analyzed by bioinformatics tools. Our results reveal information about new genes related to some cellular processes and genes involved in venom gland functions (toxins, phospholipases and antimicrobial peptides). Expressed sequence tags from venom glands provide complementary information to MS and reveal undescribed components related to the biological activity of the venom.
由电刺激尾刺(蛰针)获得了来自蝎子 Pandinus cavimanus 的毒液。总毒液对蟋蟀的毒性在 7-30μg 之间,30μg 的毒液会导致瘫痪或致死效应(每毫克蟋蟀的毒液致死 1.5μg)。电生理学分析表明,总毒液在 7μg 时对卵母细胞具有细胞溶解活性。HPLC 允许分离毒液成分。从总毒液中分离出 38 个组分,测试其对电压门控 Na(+)和 K(+)通道的影响。一些组分以不同程度抑制 K(+)电流。通过使用 MS 分析,我们从尾刺和毒液组分中获得了超过 700 种不同的分子量(通过 LC-MS/MS 和 MALDI-TOF MS 分析)。尾刺组分的二硫键数量也已确定。从 P. cavimanus 蝎子构建了 cDNA 文库,并对转录物进行了随机测序筛选。获得了不同的克隆,并通过生物信息学工具进行了分析。我们的结果揭示了与一些细胞过程相关的新基因和与毒液腺功能相关的基因(毒素、磷脂酶和抗菌肽)的信息。来自毒液腺的表达序列标签为 MS 提供了补充信息,并揭示了与毒液生物活性相关的未描述成分。