应用iTRAQ标记技术结合2DLC-MS/MS分析家蝇幼虫免疫诱导后血淋巴蛋白质组

Quantitative Proteomic Analysis on Hemolymph in Housefly Larvae with Needle Induced Using Itraq Labeling Coupled with 2D LC-MS /MS

目的分析家蝇幼虫免疫诱导前后血淋巴中的免疫防御相关靶标,探索其先天性免疫机制。方法采用同位素标记及相对定量技术(isobaric tags for relative and absolute quantification,iTRAQ)结合2D LC-MS/MS对家蝇幼虫诱导前后的血淋巴蛋白质组进行研究。结果与对照组比较研究后共获得237个不同肽段,鉴定到13个具有定量信息的差异蛋白。免疫刺激后显著上调的蛋白有9个,下调蛋白4个(P<0.05),对鉴定到的差异蛋白进行理化性质分析和GO(gene ontology)注释分析,发现这些蛋白分别具有抗菌、抗氧化、催化结合等功能,并参与了免疫、代谢、应激、转运等生物学过程,表明对家蝇幼虫经诱导后血淋巴中多种功能蛋白的表达发生了变化。结论 iTRAQ标记技术结合2D LC-MS/MS可以有效地分离鉴定昆虫血淋巴蛋白质组,为深入研究差异蛋白的功能奠定了基础。

The hemolymph of immunity induced Musca domestica larvae were studied by iTRAQ （ isobaric tags for relative and absolute quantification） coupled with 2D LC-MS/MS. When compared with the proteome in normal subjects, 237 peptides and 13 proteins were identified. The increase （decrease） numbers of proteins were 9 （4） after immunity induction （P 〈 0.05 ）. These different proteins can be functionally categorized into antimicrobial, binding, catalytic activity, and transporter activity, as well as involved in biological processes of immunity, stress, transportation and metabolism after GO analysis. It is indicated that many important functional proteins in hemolymph were expressed abnormally in the process of induction. Moreo- ver, iTRAQ labeling coupled with 2D LC-MS/MS could be successfully used for exploring the proteome of insect.