Facultad de Ciencias Químicas and Facultad de Medicina, Universidad de Colima, Colima, México.
PLoS One. 2013 Jun 20;8(6):e66486. doi: 10.1371/journal.pone.0066486. Print 2013.
Centruroides tecomanus is a Mexican scorpion endemic of the State of Colima, that causes human fatalities. This communication describes a proteome analysis obtained from milked venom and a transcriptome analysis from a cDNA library constructed from two pairs of venom glands of this scorpion. High perfomance liquid chromatography separation of soluble venom produced 80 fractions, from which at least 104 individual components were identified by mass spectrometry analysis, showing to contain molecular masses from 259 to 44,392 Da. Most of these components are within the expected molecular masses for Na(+)- and K(+)-channel specific toxic peptides, supporting the clinical findings of intoxication, when humans are stung by this scorpion. From the cDNA library 162 clones were randomly chosen, from which 130 sequences of good quality were identified and were clustered in 28 contigs containing, each, two or more expressed sequence tags (EST) and 49 singlets with only one EST. Deduced amino acid sequence analysis from 53% of the total ESTs showed that 81% (24 sequences) are similar to known toxic peptides that affect Na(+)-channel activity, and 19% (7 unique sequences) are similar to K(+)-channel especific toxins. Out of the 31 sequences, at least 8 peptides were confirmed by direct Edman degradation, using components isolated directly from the venom. The remaining 19%, 4%, 4%, 15% and 5% of the ESTs correspond respectively to proteins involved in cellular processes, antimicrobial peptides, venom components, proteins without defined function and sequences without similarity in databases. Among the cloned genes are those similar to metalloproteinases.
刺尾鳄尾蝎是一种墨西哥蝎子，仅分布于科利马州，可致人死亡。本通讯描述了从挤奶毒液中获得的蛋白质组分析和从两对毒液腺构建的 cDNA 文库中获得的转录组分析。高效液相色谱法分离可溶性毒液产生了 80 个馏分，通过质谱分析鉴定了至少 104 个单独的成分，分子量从 259 到 44,392 Da 不等。这些成分中的大多数都在预期的 Na(+)-和 K(+)-通道特异性毒性肽的分子量范围内，支持了临床中毒的发现，当人类被这种蝎子蜇伤时。从 cDNA 文库中随机选择了 162 个克隆，从中鉴定出 130 个高质量的序列，并将它们聚类成 28 个重叠群，每个重叠群包含两个或更多个表达序列标签 (EST) 和 49 个单 EST。从 53%的总 EST 中推导的氨基酸序列分析表明，81%（24 个序列）与影响 Na(+)-通道活性的已知毒性肽相似，19%（7 个独特序列）与 K(+)-通道特异性毒素相似。在 31 个序列中，至少有 8 个肽通过直接 Edman 降解得到了确认，使用的是直接从毒液中分离出的成分。其余的 19%、4%、4%、15%和 5%的 EST 分别对应于参与细胞过程的蛋白质、抗菌肽、毒液成分、无定义功能的蛋白质和无数据库相似性的序列。在克隆的基因中，有一些与金属蛋白酶相似。
