A ray of venom: Combined proteomic and transcriptomic investigation of fish venom composition using barb tissue from the blue-spotted stingray (Neotrygon kuhlii)
A ray of venom: Combined proteomic and transcriptomic investigation of fish venom composition using barb tissue from the blue-spotted stingray (Neotrygon kuhlii)
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DOI：
10.1016/j.jprot.2014.06.004
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发表时间：
2014-09-23
影响因子：
3.3
通讯作者：
Fry, Bryan G.
中科院分区：
文献类型：
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作者：
Baumann, Kate;Casewell, Nicholas R.;Fry, Bryan G.
尽管鱼毒种类繁多，但几乎完全没有人研究过。这在一定程度上是由于鱼毒的固有性质，因为它们对温度、PH值、冻干、储存和反复冻融高度敏感。它们还被粘液严重污染，这使得蛋白质组学研究变得困难。在这里，我们描述了一种新的蛋白质处理方案，利用硫酸铵和丙酮沉淀来去除粘液污染。我们使用从蓝点黄貂鱼中提取的倒钩毒腺组织蛋白来验证这种方法。我们用一维和二维凝胶对提取的蛋白质进行了LC-MS/MS测序。蛋白质注释是以毒腺转录组为基础的。我们的N.kuhlii毒液样本的组成揭示了多种蛋白质类型，这些蛋白质类型对于动物毒液系统来说是全新的。值得注意的是，检测到的蛋白质中没有一种与以前从鱼毒中发现的少数几种毒素成分相似，包括在其他黄貂鱼中发现的那些。这里确认的推定毒素包括胱抑素、过氧化还蛋白和半乳糖凝集素。我们的研究是首次对鱼类毒液器官的基因和蛋白质组成进行联合调查，我们新的蛋白质处理方法将有助于未来鉴定其他未研究的有毒鱼类的毒素。(C)2014年爱思唯尔BM。版权所有。
Fish venoms remain almost completely unstudied despite the large number of species. In part this is due to the inherent nature of fish venoms, in that they are highly sensitive to heat, pH, lyophilisation, storage and repeated freeze-thawing. They are also heavily contaminated with mucus, which makes proteomic study difficult. Here we describe a novel protein-handling protocol to remove mucus contamination, utilising ammonium sulphate and acetone precipitation. We validated this approach using barb venom gland tissue protein extract from the blue-spotted stingray Neotrygon kuhlii. We analysed the protein extract using 1D and 2D gels with LC-MS/MS sequencing. Protein annotation was underpinned by a venom gland transcriptome. The composition of our N. kuhlii venom sample revealed a variety of protein types that are completely novel to animal venom systems. Notably, none of the detected proteins exhibited similarity to the few toxin components previously characterised from fish venoms, including those found in other stingrays. Putative venom toxins identified here included cystatin, peroxiredoxin and galectin. Our study represents the first combined survey of gene and protein composition from the venom apparatus of any fish and our novel protein handling method will aid the future characterisation of toxins from other unstudied venomous fish lineages. (C) 2014 Elsevier BM. All rights reserved.