Abstract
Insect odorant receptors (ORs) have been suggested to function as ligand-gated cation channels, with OrX/Orco heteromers combining ionotropic and metabotropic activity. The latter is mediated by different G proteins and results in Orco self-activation by cyclic nucleotide binding. In this contribution, we co-express the odor-specific subunits DmOr49b and DmOr59b with either wild-type Orco or an Orco-PKC mutant lacking cAMP activation heterologously in mammalian cells. We show that the characteristics of heteromers strongly depend on both the OrX type and the coreceptor variant. Thus, methyl acetate-sensitive Or59b/Orco demonstrated 25-fold faster response kinetics over o-cresol-specific Or49b/Orco, while the latter required a 10–100 times lower ligand concentration to evoke a similar electrical response. Compared to wild-type Orco, Orco-PKC decreased odorant sensitivity in both heteromers, and blocked an outward current rectification intrinsic to the Or49b/Orco pair. Our observations thus provide an insight into insect OrX/Orco functioning, highlighting their natural and artificial tuning features and laying the groundwork for their application in chemogenetics, drug screening, and repellent design.
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Data availability
The data supporting the findings of this study are available within the manuscript and its supplementary materials. The raw data are available from the corresponding author on request.
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Acknowledgements
We sincerely thank Dr. Maksim Erokhin and Dr. Dieter Wicher for their help with the DNA templates.
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This work was supported by the Russian Foundation for Basic Research (RFBR) Grant 18-34-20087. Electrophysiology experiments were supported by Russian Science Foundation Grant 20-15-00408. L.A.K. was supported by the Russian Foundation for Basic Research (RFBR) Grant 19-34-90140 for PhD-students.
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AMB and ESN managed the project; AMB, ESN, PMB and KAL conceived and designed the experiments; DVK, LAK, and ELS performed the genetic engineering, cell culture and fluorescence microscopy experiments; VOI and ESN performed the electrophysiology measurements and analyzed the data; AMB, PMB, LAK acquired funding; AMB, VOI, ESN, and DVK wrote the paper. All authors analyzed the results and approved the final version of the manuscript.
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Kolesov, D.V., Ivanova, V.O., Sokolinskaya, E.L. et al. Impacts of OrX and cAMP-insensitive Orco to the insect olfactory heteromer activity. Mol Biol Rep 48, 4549–4561 (2021). https://doi.org/10.1007/s11033-021-06480-0
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DOI: https://doi.org/10.1007/s11033-021-06480-0