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Title: | Functional GPCR Expression in Eukaryotic LEXSY System | Authors: | Luginina, Aleksandra MASLOV, Ivan Khorn, Polina Volkov, Oleksandr Khnykin, Andrey Kuzmichev, Pavel Shevtsov, Mikhail Belousov, Anatoliy Kapranov, Ivan Dashevskii, Dmitrii Kornilov, Daniil Bestsennaia, Ekaterina Hofkens, Johan HENDRIX, Jelle Gensch, Thomas Cherezov, Vadim Ivanovich, Valentin Mishin, Alexey Borshchevskiy, Valentin |
Issue Date: | 2023 | Publisher: | ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD | Source: | JOURNAL OF MOLECULAR BIOLOGY, 435 (23) (Art N° 168310) | Abstract: | G protein-coupled receptors (GPCRs) form the largest superfamily of membrane proteins in the human genome, and represent one of the most important classes of drug targets. Their structural studies facilitate rational drug discovery. However, atomic structures of only about 20% of human GPCRs have been solved to date. Recombinant production of GPCRs for structural studies at a large scale is challenging due to their low expression levels and stability. Therefore, in this study, we explored the efficacy of the eukaryotic system LEXSY (Leishmania tarentolae) for GPCR production. We selected the human A2A adenosine receptor (A2AAR), as a model protein, expressed it in LEXSY, purified it, and compared with the same receptor produced in insect cells, which is the most popular expression system for structural studies of GPCRs. The A2AAR purified from both expression systems showed similar purity, stability, ligand-induced conformational changes and structural dynamics, with a remarkably higher protein yield in the case of LEXSY expression. Overall, our results suggest that LEXSY is a promising platform for large-scale production of GPCRs for structural studies.(c) 2023 Elsevier Ltd. All rights reserved. | Notes: | Borshchevskiy, V (corresponding author), Moscow Inst Phys & Technol, Res Ctr Mol Mech Aging & Age Related Dis, Dolgoprudnyi, Moscow, Russia.; Borshchevskiy, V (corresponding author), Joint Inst Nucl Res, Dubna, Russia. borshchevskiy.vi@phystech.edu |
Keywords: | Recombinant expression;Protein biophysical characterization;Leishmania tarentolae;Single-molecule Forster Resonance Energy Transfer;G protein-coupled receptors | Document URI: | http://hdl.handle.net/1942/42400 | ISSN: | 0022-2836 | e-ISSN: | 1089-8638 | DOI: | 10.1016/j.jmb.2023.168310 | ISI #: | 001150167700001 | Rights: | 2023 Elsevier Ltd. All rights reserved. | Category: | A1 | Type: | Journal Contribution |
Appears in Collections: | Research publications |
Files in This Item:
File | Description | Size | Format | |
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Functional GPCR Expression in Eukaryotic LEXSY System.pdf Restricted Access | Published version | 2.08 MB | Adobe PDF | View/Open Request a copy |
LEXSY_a_novel_system_for_functional_GPCR_heterologous_expression.pdf Until 2024-06-01 | Peer-reviewed author version | 1.28 MB | Adobe PDF | View/Open Request a copy |
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