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KMID : 0578320180410020150
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Molecules and Cells
2018 Volume.41 No. 2 p.150 ~ p.159
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Mass Spectrometry-Based Screening Platform Reveals Orco Interactome in Drosophila melanogaster
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Yu Kate E.
Kim Do-Hyoung
Kim Yong-In
Jones Walton D.
Lee J. Eu-Gene
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Abstract
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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.
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KEYWORD
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Drosophila, odorant receptor, Orco, IP-MS, interactome
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