Oliveira Daniele S, Brito Nathalia F, Nogueira Fabio C S, Moreira Monica F, Leal Walter S, Soares Marcia R, Melo Ana C A
Universidade Federal do Rio de Janeiro, Instituto de Química, 21941-909 Rio de Janeiro, RJ, Brazil.
Universidade Federal do Rio de Janeiro, Instituto de Química, 21941-909 Rio de Janeiro, RJ, Brazil; Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular, Rio de Janeiro, Brazil.
J Insect Physiol. 2017 Jul;100:108-118. doi: 10.1016/j.jinsphys.2017.06.004. Epub 2017 Jun 9.
Reception of odorants is essential in insects' life since the chemical signals in the environment (=semiochemicals) convey information about availability of hosts for a blood meal, mates for reproduction, sites for oviposition and other relevant information for fitness in the environment. Once they reach the antennae, these semiochemicals bind to odorant-binding proteins and are transported through the sensillar lymph until reach the odorant receptors. Such perireceptor events, particularly the interactions with transport proteins, are the liaison between the external environment and the entire neuroethological system and, therefore, a potential target to disrupt insect chemical communication. In this study, a proteomic profile of female and male antennae of Rhodnius prolixus, a vector of Chagas disease, was obtained in an attempt to unravel the entire repertoire of olfactory proteins involved in perireceptor events. Using shotgun proteomics and two-dimensional gel electrophoresis approaches followed by nano liquid chromatography coupled with tandem LTQ Velos Orbitrap mass spectrometry, we have identified 581 unique proteins. Putative olfactory proteins, including 17 odorant binding proteins, 6 chemosensory proteins, 2 odorant receptors, 3 transient receptor channels and 1 gustatory receptor were identified. Proteins involved in general cellular functions such as generation of precursor metabolites, energy generation and catabolism were expressed at high levels. Additionally, proteins that take part in signal transduction, ion binding, and stress response, kinase and oxidoreductase activity were frequent in antennae from both sexes. This proteome strategy unraveled for the first time the complex nature of perireceptor and other olfactory events that occur in R. prolixus antennae, including evidence for phosphorylation of odorant-binding and chemosensory proteins. These findings not only increase our understanding of the olfactory process in triatomine species, but also identify potential molecular targets to be explored for population control of such insect vectors.
气味分子的接收在昆虫的生命中至关重要，因为环境中的化学信号（=信息素）传达了关于宿主血餐的可获得性、繁殖配偶、产卵地点以及环境适应性的其他相关信息。一旦这些信息素到达触角，它们就会与气味结合蛋白结合，并通过感觉淋巴液运输，直到到达气味受体。这种受体周围事件，特别是与运输蛋白的相互作用，是外部环境与整个神经行为系统之间的联系，因此是破坏昆虫化学通讯的潜在靶点。在本研究中，我们获得了恰加斯病媒介昆虫——南美锥蝽雌雄触角的蛋白质组图谱，试图揭示参与受体周围事件的所有嗅觉蛋白。使用鸟枪法蛋白质组学和二维凝胶电泳方法，随后进行纳升液相色谱与串联LTQ Velos Orbitrap质谱联用，我们鉴定出了581种独特的蛋白质。鉴定出了假定的嗅觉蛋白，包括17种气味结合蛋白、6种化学感受蛋白、2种气味受体、3种瞬时受体通道和1种味觉受体。参与一般细胞功能（如前体代谢物的生成、能量产生和分解代谢）的蛋白质表达水平较高。此外，参与信号转导、离子结合、应激反应、激酶和氧化还原酶活性的蛋白质在两性触角中都很常见。这种蛋白质组策略首次揭示了南美锥蝽触角中发生的受体周围和其他嗅觉事件的复杂性质，包括气味结合蛋白和化学感受蛋白磷酸化的证据。这些发现不仅增加了我们对锥蝽物种嗅觉过程的理解，还确定了潜在的分子靶点，可用于探索控制此类昆虫媒介的种群数量。
