Laboratório de Herpetologia, Instituto Butantan, São Paulo, Brazil.
Laboratório de Herpetologia, Instituto Butantan, São Paulo, Brazil; Interunidades em Biotecnologia, Universidade de São Paulo, Instituto de Pesquisas Tecnológicas, Instituto Butantan, São Paulo, Brazil.
Toxicon. 2021 Apr 15;193:73-83. doi: 10.1016/j.toxicon.2021.01.006. Epub 2021 Jan 27.
The venom color variation of Crotalus durissus terrificus (Cdt) is attributed to the presence of the toxin L-amino acid oxidase (LAAO). During the venom milking routine of Instituto Butantan, we have noticed that most venoms of captive Cdt specimens show a yellowish color, while most venoms of wild specimens are white. Here we describe a comparative analysis of long-term captive (LTC) and recently wild-caught (RWC) Cdt, focusing on LAAO variation. For the identification of LAAO in individual venoms, four different approaches were employed: evaluation of the enzymatic activity, SDS-PAGE, Western blotting, and ELISA. In addition, mass spectrometry analysis was performed using pooled samples. Although some variation among these methodologies was observed, it was possible to notice that the presence of LAAO was significantly higher in the venom of LTC individuals. LAAO was identified in 60-80% LTC specimens and in only 10-12% of RWC specimens. Furthermore, this enzyme accounts for 5.6% of total venom proteins of LTC Cdt pooled venom, while it corresponds to only 0.7% of RWC Cdt pooled venom. These findings strongly suggest that captive maintenance increases the expression of LAAO in Cdt venom.
响尾蛇属(Crotalus durissus terrificus)毒液颜色的变化归因于毒素 L-氨基酸氧化酶(LAAO)的存在。在布坦坦研究所的毒液采集常规操作中,我们注意到大多数圈养响尾蛇标本的毒液呈黄色,而大多数野生标本的毒液呈白色。在这里,我们描述了对长期圈养(LTC)和最近野外捕获(RWC)响尾蛇的比较分析,重点是 LAAO 的变化。为了鉴定个体毒液中的 LAAO,我们采用了四种不同的方法:酶活性评估、SDS-PAGE、Western blot 和 ELISA。此外,还使用混合样本进行了质谱分析。尽管在这些方法学之间观察到了一些差异,但可以注意到,LAAO 在 LTC 个体的毒液中的存在显著更高。LAAO 在 60-80%的 LTC 标本中被鉴定出来,而在 10-12%的 RWC 标本中被鉴定出来。此外,这种酶占 LTC 响尾蛇混合毒液总毒液蛋白的 5.6%,而 RWC 响尾蛇混合毒液中仅占 0.7%。这些发现强烈表明,圈养饲养会增加响尾蛇毒液中 LAAO 的表达。