Ligand-directed covalent labelling of a GPCR with a fluorescent tag in live cells.


Journal

Communications biology
ISSN: 2399-3642
Titre abrégé: Commun Biol
Pays: England
ID NLM: 101719179

Informations de publication

Date de publication:
27 11 2020
Historique:
received: 15 05 2020
accepted: 03 11 2020
entrez: 28 11 2020
pubmed: 29 11 2020
medline: 24 6 2021
Statut: epublish

Résumé

To study the localisation of G protein-coupled receptors (GPCR) in their native cellular environment requires their visualisation through fluorescent labelling. To overcome the requirement for genetic modification of the receptor or the limitations of dissociable fluorescent ligands, here we describe rational design of a compound that covalently and selectively labels a GPCR in living cells with a fluorescent moiety. We designed a fluorescent antagonist, in which the linker incorporated between pharmacophore (ZM241385) and fluorophore (sulfo-cyanine5) is able to facilitate covalent linking of the fluorophore to the adenosine A

Identifiants

pubmed: 33247190
doi: 10.1038/s42003-020-01451-w
pii: 10.1038/s42003-020-01451-w
pmc: PMC7695831
doi:

Substances chimiques

ADORA2A protein, human 0
Affinity Labels 0
Fluorescent Dyes 0
Ligands 0
Receptor, Adenosine A2A 0
Receptors, G-Protein-Coupled 0
Triazines 0
Triazoles 0
ZM 241385 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

722

Subventions

Organisme : Medical Research Council
ID : MR/N020081/1
Pays : United Kingdom
Organisme : Department of Health
Pays : United Kingdom

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Auteurs

Leigh A Stoddart (LA)

Cell Signalling and Pharmacology Research Group, Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham, NG7 2RD, UK.
Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Nottingham, Midlands, NG7 2RD, UK.

Nicholas D Kindon (ND)

Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Nottingham, Midlands, NG7 2RD, UK.
School of Pharmacy, Biodiscovery Institute, University of Nottingham, Nottingham, NG7 2RD, UK.

Omolade Otun (O)

Cell Signalling and Pharmacology Research Group, Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham, NG7 2RD, UK.
Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Nottingham, Midlands, NG7 2RD, UK.

Clare R Harwood (CR)

Cell Signalling and Pharmacology Research Group, Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham, NG7 2RD, UK.
Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Nottingham, Midlands, NG7 2RD, UK.

Foteini Patera (F)

Division of Cancer and Stem Cells, School of Medicine, University of Nottingham, Nottingham, NG7 2RD, UK.

Dmitry B Veprintsev (DB)

Cell Signalling and Pharmacology Research Group, Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham, NG7 2RD, UK.
Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Nottingham, Midlands, NG7 2RD, UK.

Jeanette Woolard (J)

Cell Signalling and Pharmacology Research Group, Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham, NG7 2RD, UK.
Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Nottingham, Midlands, NG7 2RD, UK.

Stephen J Briddon (SJ)

Cell Signalling and Pharmacology Research Group, Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham, NG7 2RD, UK.
Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Nottingham, Midlands, NG7 2RD, UK.

Hester A Franks (HA)

Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Nottingham, Midlands, NG7 2RD, UK.
Division of Cancer and Stem Cells, School of Medicine, University of Nottingham, Nottingham, NG7 2RD, UK.

Stephen J Hill (SJ)

Cell Signalling and Pharmacology Research Group, Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham, NG7 2RD, UK. stephen.hill@nottingham.ac.uk.
Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Nottingham, Midlands, NG7 2RD, UK. stephen.hill@nottingham.ac.uk.

Barrie Kellam (B)

Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Nottingham, Midlands, NG7 2RD, UK. barrie.kellam@nottingham.ac.uk.
School of Pharmacy, Biodiscovery Institute, University of Nottingham, Nottingham, NG7 2RD, UK. barrie.kellam@nottingham.ac.uk.

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Classifications MeSH