Himaya S W A, Jin Ai-Hua, Dutertre Sébastien, Giacomotto Jean, Mohialdeen Hoshyar, Vetter Irina, Alewood Paul F, Lewis Richard J
Institute for Molecular Bioscience, The University of Queensland , Brisbane, 4072 Queensland, Australia.
Institut des Biomolécules Max Mousseron, UMR 5247, Université Montpellier-CNRS , Place Eugène Bataillon, Montpellier Cedex 5 34095, France.
J Proteome Res. 2015 Oct 2;14(10):4372-81. doi: 10.1021/acs.jproteome.5b00630. Epub 2015 Sep 10.
Venomous marine cone snails produce a unique and remarkably diverse range of venom peptides (conotoxins and conopeptides) that have proven to be invaluable as pharmacological probes and leads to new therapies. Conus catus is a hook-and-line fish hunter from clade I, with ∼20 conotoxins identified, including the analgesic ω-conotoxin CVID (AM336). The current study unravels the venom composition of C. catus with tandem mass spectrometry and 454 sequencing data. From the venom gland transcriptome, 104 precursors were recovered from 11 superfamilies, with superfamily A (especially κA-) conotoxins dominating (77%) their venom. Proteomic analysis confirmed that κA-conotoxins dominated the predation-evoked milked venom of each of six C. catus analyzed and revealed remarkable intraspecific variation in both the intensity and type of conotoxins. High-throughput FLIPR assays revealed that the predation-evoked venom contained a range of conotoxins targeting the nAChR, Cav, and Nav ion channels, consistent with α- and ω-conotoxins being used for predation by C. catus. However, the κA-conotoxins did not act at these targets but induced potent and rapid immobilization followed by bursts of activity and finally paralysis when injected intramuscularly in zebrafish. Our venomics approach revealed the complexity of the envenomation strategy used by C. catus, which contains a mix of both excitatory and inhibitory venom peptides.
有毒的海洋芋螺会产生一系列独特且极为多样的毒液肽(芋螺毒素和芋螺缩氨酸),这些毒液肽已被证明作为药理学探针具有极高价值,并能为新疗法提供线索。Catus芋螺是来自进化枝I的一种用钩线捕鱼的猎手,已鉴定出约20种芋螺毒素,包括止痛的ω-芋螺毒素CVID(AM336)。当前的研究通过串联质谱和454测序数据揭示了Catus芋螺的毒液成分。从毒液腺转录组中,从11个超家族中回收了104个前体,其中超家族A(尤其是κA-)芋螺毒素占其毒液的主导地位(77%)。蛋白质组学分析证实,κA-芋螺毒素在分析的6只Catus芋螺的每一只的捕食诱发的挤取毒液中占主导地位,并揭示了芋螺毒素在强度和类型上都存在显著的种内变异。高通量FLIPR分析表明,捕食诱发的毒液含有一系列靶向nAChR、Cav和Nav离子通道的芋螺毒素,这与α-和ω-芋螺毒素被Catus芋螺用于捕食一致。然而,κA-芋螺毒素并不作用于这些靶点,而是在肌肉注射到斑马鱼体内时诱导出强力且快速的固定,随后是一阵活动,最终导致麻痹。我们的毒液组学方法揭示了Catus芋螺所采用的中毒策略的复杂性,其毒液中含有兴奋性和抑制性毒液肽的混合物。