Mol. Cells 2018; 41(2): 150~159  https://doi.org/10.14348/molcells.2018.2305
Mass Spectrometry-Based Screening Platform Reveals Orco Interactome in Drosophila melanogaster
Kate E. Yu1,2, Do-Hyoung Kim1,3, Yong-In Kim2, Walton D. Jones1,*, and J. Eugene Lee2,*
1Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon 34141, Korea, 2Center for
Bioanalysis, Korea Research Institute of Standards and Science, Daejeon 34113, Korea, 3Present address: School of Life Sciences,
Gwangju Institute of Science and Technology, Gwangju 61005, Korea
*Correspondence: waltonjones@kaist.edu (WJ); j.eugenelee@gmail.com (JL)
Received November 17, 2017; Revised December 11, 2017; Accepted December 12, 2017.; Published online February 12, 2018.
© Korean Society for Molecular and Cellular Biology. All rights reserved.

This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License. To view a copy of this license, visit (http://creativecommons.org/licenses/by-nc-sa/3.0/).
ABSTRACT
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.
Keywords: Drosophila, odorant receptor, Orco, IP-MS, interactome


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