Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/42400
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dc.contributor.authorLuginina, Aleksandra-
dc.contributor.authorMASLOV, Ivan-
dc.contributor.authorKhorn, Polina-
dc.contributor.authorVolkov, Oleksandr-
dc.contributor.authorKhnykin, Andrey-
dc.contributor.authorKuzmichev, Pavel-
dc.contributor.authorShevtsov, Mikhail-
dc.contributor.authorBelousov, Anatoliy-
dc.contributor.authorKapranov, Ivan-
dc.contributor.authorDashevskii, Dmitrii-
dc.contributor.authorKornilov, Daniil-
dc.contributor.authorBestsennaia, Ekaterina-
dc.contributor.authorHofkens, Johan-
dc.contributor.authorHENDRIX, Jelle-
dc.contributor.authorGensch, Thomas-
dc.contributor.authorCherezov, Vadim-
dc.contributor.authorIvanovich, Valentin-
dc.contributor.authorMishin, Alexey-
dc.contributor.authorBorshchevskiy, Valentin-
dc.date.accessioned2024-02-13T11:02:56Z-
dc.date.available2024-02-13T11:02:56Z-
dc.date.issued2023-
dc.date.submitted2024-02-13T09:58:59Z-
dc.identifier.citationJOURNAL OF MOLECULAR BIOLOGY, 435 (23) (Art N° 168310)-
dc.identifier.urihttp://hdl.handle.net/1942/42400-
dc.description.abstractG 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.-
dc.description.sponsorshipAuthors thank Taras Balandin for the advising and discussion during project realization. This work is supported by RSCF research grant 22-74-00024 (https://rscf.ru/en/project/22-74-0002 4/). J. Ho. acknowledges support from the Flemish government through long-term structural funding Methusalem (CASAS2, Meth/15/04). J. He acknowledges the Research Foundation Flanders (FWO grants G0B9922N).-
dc.language.isoen-
dc.publisherACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD-
dc.rights2023 Elsevier Ltd. All rights reserved.-
dc.subject.otherRecombinant expression-
dc.subject.otherProtein biophysical characterization-
dc.subject.otherLeishmania tarentolae-
dc.subject.otherSingle-molecule Forster Resonance Energy Transfer-
dc.subject.otherG protein-coupled receptors-
dc.titleFunctional GPCR Expression in Eukaryotic LEXSY System-
dc.typeJournal Contribution-
dc.identifier.issue23-
dc.identifier.volume435-
local.format.pages16-
local.bibliographicCitation.jcatA1-
dc.description.notesBorshchevskiy, 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.-
dc.description.notesborshchevskiy.vi@phystech.edu-
local.publisher.place24-28 OVAL RD, LONDON NW1 7DX, ENGLAND-
local.type.refereedRefereed-
local.type.specifiedArticle-
local.bibliographicCitation.artnr168310-
dc.identifier.doi10.1016/j.jmb.2023.168310-
dc.identifier.pmid37806553-
dc.identifier.isi001150167700001-
dc.contributor.orcidLuginina, Aleksandra/0000-0003-2697-456X; Shevtsov,-
dc.contributor.orcidMikhail/0000-0001-8160-2967; Andrey, Khnykin/0009-0006-2631-013X;-
dc.contributor.orcidMaslov, Ivan/0000-0003-3371-4416; Khorn, Polina/0000-0002-2117-2130;-
dc.contributor.orcidBelousov, Anatoliy/0009-0008-4479-1517; Dashevskii,-
dc.contributor.orcidDmitrii/0000-0001-9901-6704; Kuzmichev, Pavel/0000-0001-5322-3580-
local.provider.typewosris-
local.description.affiliation[Luginina, Aleksandra; Maslov, Ivan; Khorn, Polina; Khnykin, Andrey; Kuzmichev, Pavel; Shevtsov, Mikhail; Belousov, Anatoliy; Kapranov, Ivan; Dashevskii, Dmitrii; Kornilov, Daniil; Bestsennaia, Ekaterina; Ivanovich, Valentin; Mishin, Alexey; Borshchevskiy, Valentin] Moscow Inst Phys & Technol, Res Ctr Mol Mech Aging & Age Related Dis, Dolgoprudnyi, Moscow, Russia.-
local.description.affiliation[Maslov, Ivan; Hendrix, Jelle] Hasselt Univ, Biomed Res Inst, Adv Opt Microscopy Ctr, Agoralaan BIOMED C,Dynam Bioimaging Lab, Diepenbeek, Belgium.-
local.description.affiliation[Maslov, Ivan; Hofkens, Johan; Hendrix, Jelle; Gensch, Thomas] Katholieke Univ Leuven, Dept Chem, Div Mol Imaging & Photon, Lab Photochem & Spect, Leuven, Belgium.-
local.description.affiliation[Hofkens, Johan] Max Planck Inst Polymer Res, Mainz, Germany.-
local.description.affiliation[Cherezov, Vadim] Univ Southern Calif, Bridge Inst, Dept Chem, Los Angeles, CA USA.-
local.description.affiliation[Borshchevskiy, Valentin] Joint Inst Nucl Res, Dubna, Russia.-
local.uhasselt.internationalyes-
item.contributorLuginina, Aleksandra-
item.contributorMASLOV, Ivan-
item.contributorKhorn, Polina-
item.contributorVolkov, Oleksandr-
item.contributorKhnykin, Andrey-
item.contributorKuzmichev, Pavel-
item.contributorShevtsov, Mikhail-
item.contributorBelousov, Anatoliy-
item.contributorKapranov, Ivan-
item.contributorDashevskii, Dmitrii-
item.contributorKornilov, Daniil-
item.contributorBestsennaia, Ekaterina-
item.contributorHofkens, Johan-
item.contributorHENDRIX, Jelle-
item.contributorGensch, Thomas-
item.contributorCherezov, Vadim-
item.contributorIvanovich, Valentin-
item.contributorMishin, Alexey-
item.contributorBorshchevskiy, Valentin-
item.accessRightsEmbargoed Access-
item.fulltextWith Fulltext-
item.fullcitationLuginina, 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 (2023) Functional GPCR Expression in Eukaryotic LEXSY System. In: JOURNAL OF MOLECULAR BIOLOGY, 435 (23) (Art N° 168310).-
item.embargoEndDate2024-06-01-
crisitem.journal.issn0022-2836-
crisitem.journal.eissn1089-8638-
Appears in Collections:Research publications
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