Laboratory of Zoophysiology, Ghent University, Ghent, Belgium.
Insect Mol Biol. 2010 Feb;19 Suppl 1(Suppl 1):11-26. doi: 10.1111/j.1365-2583.2009.00914.x.
With the Nasonia vitripennis genome sequences available, we attempted to determine the proteins present in venom by two different approaches. First, we searched for the transcripts of venom proteins by a bioinformatic approach using amino acid sequences of known hymenopteran venom proteins. Second, we performed proteomic analyses of crude N. vitripennis venom removed from the venom reservoir, implementing both an off-line two-dimensional liquid chromatography matrix-assisted laser desorption/ ionization time-of-flight (2D-LC-MALDI-TOF) mass spectrometry (MS) and a two-dimensional liquid chromatography electrospray ionization Founer transform ion cyclotron resonance (2D-LC-ESI-FT-ICR) MS setup. This combination of bioinformatic and proteomic studies resulted in an extraordinary richness of identified venom constituents. Moreover, half of the 79 identified proteins were not yet associated with insect venoms: 16 proteins showed similarity only to known proteins from other tissues or secretions, and an additional 23 did not show similarity to any known protein. Serine proteases and their inhibitors were the most represented. Fifteen nonsecretory proteins were also identified by proteomic means and probably represent so-called 'venom trace elements'. The present study contributes greatly to the understanding of the biological diversity of the venom of parasitoid wasps at the molecular level.
有了 Nasonia vitripennis 的基因组序列,我们尝试通过两种不同的方法来确定毒液中的蛋白质。首先,我们使用已知膜翅目毒液蛋白的氨基酸序列,通过生物信息学方法搜索毒液蛋白的转录本。其次,我们对从毒液库中提取的粗 N. vitripennis 毒液进行了蛋白质组学分析,同时采用离线二维液相色谱 - 基质辅助激光解吸/电离飞行时间(2D-LC-MALDI-TOF)质谱(MS)和二维液相色谱电喷雾电离傅里叶变换离子回旋共振(2D-LC-ESI-FT-ICR)MS 两种方法。这种生物信息学和蛋白质组学研究的结合产生了异常丰富的鉴定毒液成分。此外,在鉴定出的 79 种蛋白质中,有一半以前并未与昆虫毒液有关联:16 种蛋白质仅与其他组织或分泌物中的已知蛋白质具有相似性,还有 23 种蛋白质与任何已知蛋白质均无相似性。丝氨酸蛋白酶及其抑制剂是最主要的成分。通过蛋白质组学方法还鉴定出 15 种非分泌蛋白,它们可能代表所谓的“毒液微量元素”。本研究在分子水平上极大地促进了对寄生蜂毒液生物学多样性的理解。