School of Life Sciences, University of Technology Sydney, Broadway, NSW 2007, Australia.
CNRS, UMR Ecologie des Forêts de Guyane, AgroParisTech, CIRAD, INRA, Université de Guyane, Université des Antilles, Campus Agronomique, BP316, CEDEX, 97379 Kourou, France.
Toxins (Basel). 2020 May 14;12(5):324. doi: 10.3390/toxins12050324.
A critical hurdle in ant venom proteomic investigations is the lack of databases to comprehensively and specifically identify the sequence and function of venom proteins and peptides. To resolve this, we used venom gland transcriptomics to generate a sequence database that was used to assign the tandem mass spectrometry (MS) fragmentation spectra of venom peptides and proteins to specific transcripts. This was performed alongside a shotgun liquid chromatography-mass spectrometry (LC-MS/MS) analysis of the venom to confirm that these assigned transcripts were expressed as proteins. Through the combined transcriptomic and proteomic investigation of venom, we identified four times the number of proteins previously identified using 2D-PAGE alone. In addition to this, by mining the transcriptomic data, we identified several novel peptide sequences for future pharmacological investigations, some of which conform with inhibitor cysteine knot motifs. These types of peptides have the potential to be developed into pharmaceutical or bioinsecticide peptides.
在抗毒液蛋白质组学研究中,一个关键的障碍是缺乏数据库来全面、专门地识别毒液蛋白质和肽的序列和功能。为了解决这个问题,我们使用毒液腺转录组学生成了一个序列数据库,该数据库用于将毒液肽和蛋白质的串联质谱(MS)碎片谱分配给特定的转录本。同时,我们还对毒液进行了鸟枪法液相色谱-质谱(LC-MS/MS)分析,以确认这些分配的转录本是否表达为蛋白质。通过对毒液的转录组学和蛋白质组学联合研究,我们鉴定出的蛋白质数量是以前仅使用 2D-PAGE 鉴定出的数量的四倍。此外,通过挖掘转录组数据,我们鉴定出了一些新的肽序列,用于未来的药理学研究,其中一些符合抑制剂半胱氨酸结基序。这些类型的肽有可能被开发成药物或生物杀虫剂肽。