Santibáñez-López Carlos E, Cid-Uribe Jimena I, Zamudio Fernando Z, Batista Cesar V F, Ortiz Ernesto, Possani Lourival D
Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Avenida Universidad 2001, Apartado Postal 510-3, Cuernavaca, Morelos, 62210, Mexico.
Laboratorio Universitario de Proteómica, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Avenida Universidad 2001, Apartado Postal 510-3, Cuernavaca, Morelos, 62210, Mexico.
Toxicon. 2017 Jul;133:95-109. doi: 10.1016/j.toxicon.2017.05.002. Epub 2017 May 3.
The soluble venom from the Mexican scorpion Megacormus gertschi of the family Euscorpiidae was obtained and its biological effects were tested in several animal models. This venom is not toxic to mice at doses of 100 μg per 20 g of mouse weight, while being lethal to arthropods (insects and crustaceans), at doses of 20 μg (for crickets) and 100 μg (for shrimps) per animal. Samples of the venom were separated by high performance liquid chromatography and circa 80 distinct chromatographic fractions were obtained from which 67 components have had their molecular weights determined by mass spectrometry analysis. The N-terminal amino acid sequence of seven protein/peptides were obtained by Edman degradation and are reported. Among the high molecular weight components there are enzymes with experimentally-confirmed phospholipase activity. A pair of telsons from this scorpion species was dissected, from which total RNA was extracted and used for cDNA library construction. Massive sequencing by the Illumina protocol, followed by de novo assembly, resulted in a total of 110,528 transcripts. From those, we were able to annotate 182, which putatively code for peptides/proteins with sequence similarity to previously-reported venom components available from different protein databases. Transcripts seemingly coding for enzymes showed the richest diversity, with 52 sequences putatively coding for proteases, 20 for phospholipases, 8 for lipases and 5 for hyaluronidases. The number of different transcripts potentially coding for peptides with sequence similarity to those that affect ion channels was 19, for putative antimicrobial peptides 19, and for protease inhibitor-like peptides, 18. Transcripts seemingly coding for other venom components were identified and described. The LC/MS analysis of a trypsin-digested venom aliquot resulted in 23 matches with the translated transcriptome database, which validates the transcriptome. The proteomic and transcriptomic analyses reported here constitute the first approach to study the venom components from a scorpion species belonging to the family Euscorpiidae. The data certainly show that this venom is different from all the ones described thus far in the literature.
从真蝎科的墨西哥蝎子Megacormus gertschi中获取了可溶性毒液，并在几种动物模型中测试了其生物学效应。该毒液对体重20克的小鼠以每克100微克的剂量无毒，而对节肢动物（昆虫和甲壳类动物）则具有致死性，对每只动物的剂量分别为20微克（对蟋蟀）和100微克（对虾）。毒液样品通过高效液相色谱法进行分离，获得了大约80个不同的色谱馏分，其中67个组分的分子量通过质谱分析确定。通过埃德曼降解法获得了7种蛋白质/肽的N端氨基酸序列并进行了报道。在高分子量组分中，有经实验证实具有磷脂酶活性的酶。解剖了一对该蝎子物种的尾节，从中提取了总RNA并用于构建cDNA文库。通过Illumina协议进行大规模测序，然后进行从头组装，总共产生了110,528个转录本。从中，我们能够注释182个，这些转录本推测编码与不同蛋白质数据库中先前报道的毒液成分具有序列相似性的肽/蛋白质。似乎编码酶的转录本显示出最丰富的多样性，有52个序列推测编码蛋白酶，20个编码磷脂酶，8个编码脂肪酶，5个编码透明质酸酶。可能编码与影响离子通道的肽具有序列相似性的肽的不同转录本数量为19个，推测抗菌肽为19个，蛋白酶抑制剂样肽为18个。鉴定并描述了似乎编码其他毒液成分的转录本。对胰蛋白酶消化的毒液等分试样进行的LC/MS分析与翻译后的转录组数据库产生了23个匹配，这验证了转录组。本文报道的蛋白质组学和转录组学分析构成了研究真蝎科蝎子物种毒液成分的首次方法。数据确实表明，这种毒液与文献中迄今为止描述的所有毒液都不同。
