【专集论文】Acrostyle的蛋白质组组成:鉴定参与病毒–昆虫互作的表皮蛋白的新方法

2023-05-10 14:56:27

Acrostyle是蚜虫下颚针顶端普遍存在的食物/唾液道内表面表皮上的一个明显的解剖部位。这一保守结构引起了人们特别的兴趣,原因是它至少含有一种植物病毒即黄瓜花叶病毒Cucumbermosaic virus的蛋白受体,推测其在植物昆虫互作中还发挥着其他作用。之前我们报道了对acrostyleCRR家族(因存在RebersRiddiford共有序列而命名)表皮蛋白的一个高度保守基序进行的免疫标记。

    本研究报道了应用新手段来进一步研究这一区域的蛋白组组成,鉴定参与昆虫病毒互作的蛋白。应用抗PR-2亚家族表皮蛋白的一系列抗体,鉴定了acrostyle内存在的其他肽。结果表明,acrostyle是一个复杂的结构,含有可进行互作的表皮蛋白的多个结构域。此外,建立了体现PR-2亚家族大部分成员多样性的具有重复序列的多肽阵列,作为遗传工具,来鉴定表皮蛋白病原菌之间的互作。用这一阵列探测黄瓜花叶病毒粒子,结果从杂交的肽段中鉴定到了共有肽序列。应用这些新手段,拓展了昆虫下颚针蛋白组组成方面的知识,鉴定了参与病毒结合的序列,从而可进一步阐明acrostyle的不同性质和功能。

Fig. 1 Mapping of accessible peptides of the RR-2 chitin-binding domain within the acrostyle of Acyrthosiphon pisum by immunolabeling with peptide-derived antisera. (A) Schematic representation of the location of peptides used for antibodies production. A typical RR-2 cuticular protein is represented, with the signal peptide (SP), and the N-terminal and C-terminal domains surrounding the central RR-2 chitin-binding domain. The peptides sequences (black andorange lines) are targeted upstream and downstream of the peptide pepL of the RR-2 domain. Peptides likely to be accessible within the acrostyle identified by immunolabeling of dissected individualized stylets are in bold. (B) Immunolabeling of A. pisum stylets showing variety of labeling seen with stylets both undigested (untreated, top row) and digested for 15 min with chitinase (chitinase, bottom row). Scale bars of 5 μm are included. Immunolabeling is shown for anti-pepS, anti-pepL, anti-Ap2- 05, anti-Ap2-08, and anti-Ap2-09 antibodies.


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Webster, C.G., Thillier, M., Pirolles, E., Cayrol, B., Blanc, S. and Uzest, M. (2017) Proteomic composition of the acrostyle: Novel approaches to identify cuticular proteins involved in virus–insect interactions. Insect Science, 24(6), 990–1002.

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