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. 2022 Oct 21;12:17658. doi: 10.1038/s41598-022-21678-3

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Screening of signal sequences to increase functional expression of TAS2R20/38/50 at the plasma membrane. (a) Schematic of the design and principles of the cell surface expression HiBiT assay. (b) Heat map of the fold increase in luminescence signals of TAS2R20/38/50 tagged with various N-terminal signal sequences compared with untransfected cells. The signal sequence derived from the M₃ receptor, indicated by a red circle, generated the highest surface TAS2R expression for TAS2R20/38/50. (c) Concentration–response curves of TAS2R20/38/50 upon stimulation with their cognate agonists in the bioluminescence-based intracellular calcium release assay. M₃-TAS2Rs had higher overall magnitude of responses compared to SST₃-TAS2Rs. The identities of GPCRs used for screening are abbreviated according to the nomenclature employed by the IUPHAR/BPS Concise Guide to Pharmacology (http://www.guidetopharmacology.org). Adrenergic receptor is abbreviated to AR for simplicity. Data points are shown as mean ± s.e.m. from a representative experiment out of three independent biological replicates performed in technical quadruplicates. EC₅₀, half-maximal effective concentration. (a) and (b) were generated using Adobe Illustrator (Version 25.4.1) and Prism Mac (Version 7.0e), respectively.