Institute for Molecular Bioscience, University of Queensland, St. Lucia, QLD 4072, Australia.
Centre for Advanced Imaging, University of Queensland, St. Lucia, QLD 4072, Australia.
J Proteomics. 2017 Aug 23;166:83-92. doi: 10.1016/j.jprot.2017.07.007. Epub 2017 Jul 21.
More than a century of research on sea anemone venoms has shown that they contain a diversity of biologically active proteins and peptides. However, recent omics studies have revealed that much of the venom proteome remains unexplored. We used, for the first time, a combination of proteomic and transcriptomic techniques to obtain a holistic overview of the venom arsenal of the well-studied sea anemone Stichodactyla haddoni. A purely search-based approach to identify putative toxins in a transcriptome from tentacles regenerating after venom extraction identified 508 unique toxin-like transcripts grouped into 63 families. However, proteomic analysis of venom revealed that 52 of these toxin families are likely false positives. In contrast, the combination of transcriptomic and proteomic data enabled positive identification of 23 families of putative toxins, 12 of which have no homology known proteins or peptides. Our data highlight the importance of using proteomics of milked venom to correctly identify venom proteins/peptides, both known and novel, while minimizing false positive identifications from non-toxin homologues identified in transcriptomes of venom-producing tissues. This work lays the foundation for uncovering the role of individual toxins in sea anemone venom and how they contribute to the envenomation of prey, predators, and competitors.
Proteomic analysis of milked venom combined with analysis of a tentacle transcriptome revealed the full extent of the venom arsenal of the sea anemone Stichodactyla haddoni. This combined approach led to the discovery of 12 entirely new families of disulfide-rich peptides and proteins in a genus of anemones that have been studied for over a century.
一个多世纪以来对海葵毒液的研究表明,它们含有多种具有生物活性的蛋白质和肽。然而,最近的组学研究表明,大部分毒液蛋白质组仍未被探索。我们首次使用蛋白质组学和转录组学技术的组合,全面了解了研究充分的海葵石珊瑚毒液库。在从毒液提取后再生的触手中,基于搜索的方法识别出 508 种独特的类毒素转录本,这些转录本分为 63 个家族。然而,毒液的蛋白质组学分析表明,其中 52 个毒素家族可能是假阳性。相比之下,转录组学和蛋白质组学数据的组合使我们能够确定 23 个假定毒素家族,其中 12 个家族没有已知蛋白质或肽的同源性。我们的数据强调了使用挤奶毒液的蛋白质组学来正确识别毒液蛋白/肽的重要性,包括已知和新的蛋白/肽,同时最大限度地减少从产毒液组织的转录本中鉴定出的非毒素同源物的假阳性鉴定。这项工作为揭示单个毒素在海葵毒液中的作用以及它们如何导致猎物、捕食者和竞争者中毒奠定了基础。
挤奶毒液的蛋白质组学分析与触须转录组分析相结合,揭示了海葵石珊瑚毒液库的全部内容。这种组合方法在一个已经研究了一个多世纪的海葵属中发现了 12 种全新的富含二硫键的肽和蛋白质家族。