Abstract

The venom of Lycosoidea spiders is a complex multicomponent mixture of neurotoxic peptides (main components) and antimicrobial peptides (AMPs) as minor components. In this study, we described the high-throughput identification and analysis of AMPs from Lycosa sinensis venom (named LS-AMPs) using a combination strategy that includes the following three different analysis approaches: (i) peptidomic analysis, namely reversed-phase high-performance liquid chromatography (RP-HPLC) separation plus top-down sequencing by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MS); (ii) transcriptomic analysis, namely cDNA library construction plus DNA sequencing; (iii) bioinformatic analysis, namely analysis and prediction for molecular characters of LS-AMPs by the online biology databases. In total, 52 sequences of AMPs were identified from L. sinensis venom, and all AMPs can be categorized into eight different families according to phylogenetic analysis and sequence identity. This is the largest number of AMPs identified from a spider species so far. In the present study, we demonstrated molecular characteristics, such as complex precursor, N- and/or C-terminally truncated analogs, and C-terminal amidation of LS-AMPs from L. sinensis venom. This is a preliminary investigation on the molecular diversification of venom-derived AMPs from the wolf spider species (family Lycosidae), and a detailed investigation on the functional diversity of LS-AMPs will be preformed in the future.

中文翻译：

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基于肽组学，转录组学和生物信息学分析的狼蛛Lycosa sinensis毒液中抗菌肽的分子多样性

摘要

Lycosoidea蜘蛛的毒液是神经毒性肽（主要成分）和抗菌肽（AMPs）的次要成分的复杂多成分混合物。在这项研究中，我们描述了放大器的高通量鉴定和分析来自狼蛛冬虫夏草毒液（称为LS-AMPs），其使用包括以下三种不同分析方法的组合策略：（i）肽组分析，即反相高效液相色谱（RP-HPLC）分离加上通过基质辅助的自顶向下测序激光解吸/电离飞行时间质谱（MS）; （ii）转录组分析，即cDNA文库构建和DNA测序；（iii）生物信息学分析，即通过在线生物学数据库对LS-AMPs的分子特征进行分析和预测。总共从中华乳杆菌中鉴定出52个AMP序列毒液，根据系统发育分析和序列同一性，所有AMP均可分为八个不同的家族。到目前为止，这是从蜘蛛物种中鉴定出的最大数量的AMP。在本研究中，我们证明了分子特征，例如复杂的前体，N和/或C端截短的类似物以及L. sinensis毒液中LS-AMP的C端酰胺化。这是对狼蛛物种（Lycosidae科）中毒液衍生的AMPs分子多样性的初步研究，将来将对LS-AMPs的功能多样性进行详细的研究。