Mass Spectrometry-Based Screening Platform Reveals Orco Interactome in Drosophila melanogaster.
Mass Spectrometry-Based Screening Platform Reveals Orco Interactome in Drosophila melanogaster.
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DOI：
10.14348/molcells.2018.2305
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发表时间：
2018-02-28
影响因子：
3.8
通讯作者：
Lee JE
中科院分区：
文献类型：
--
作者：
Yu KE;Kim DH;Kim YI;Jones WD;Lee JE
动物使用它们的气味受体来接收来自环境的化学信息。昆虫气味受体不同于脊椎动物和线虫中的G蛋白偶联气味受体，并且对它们的蛋白质-蛋白质相互作用知之甚少。在这里，我们介绍了一个质谱平台，设计用于大规模分析昆虫气味受体蛋白质-蛋白质相互作用。利用该平台，我们从果蝇中获得了第一个Orco相互作用组。从总共1，186个已鉴定的蛋白质中，我们将相互作用候选者缩小到226个，其中只有三分之二被命名。这些候选者包括已知的嗅觉蛋白Or 92 a和Obp 51 a。大约90%的蛋白质可能在细胞内起作用，这些细胞内蛋白质中有近一半与内膜系统有关。在一个基本的功能丧失电生理学筛选中，我们发现八个（即，Rab 5、CG 32795、Mpcp、Tom 70、Vir-1、CG 30427、Eaat 1和CG 2781）影响体内嗅觉反应。因此，由于这个Orco相互作用组包括具有生理意义的候选物，我们预计我们的平台将有助于指导对昆虫气味受体家族分子机制的进一步研究。
Animals use their odorant receptors to receive chemical information from the environment. Insect odorant receptors differ from the G protein-coupled odorant receptors in vertebrates and nematodes, and very little is known about their protein–protein interactions. Here, we introduce a mass spectrometric platform designed for the large-scale analysis of insect odorant receptor protein–protein interactions. Using this platform, we obtained the first Orco interactome from Drosophila melanogaster. From a total of 1,186 identified proteins, we narrowed the interaction candidates to 226, of which only two-thirds have been named. These candidates include the known olfactory proteins Or92a and Obp51a. Around 90% of the proteins having published names likely function inside the cell, and nearly half of these intracellular proteins are associated with the endomembrane system. In a basic loss-of-function electrophysiological screen, we found that the disruption of eight (i.e., Rab5, CG32795, Mpcp, Tom70, Vir-1, CG30427, Eaat1, and CG2781) of 28 randomly selected candidates affects olfactory responses in vivo. Thus, because this Orco interactome includes physiologically meaningful candidates, we anticipate that our platform will help guide further research on the molecular mechanisms of the insect odorant receptor family.