Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

Ligand-directed covalent labelling of a GPCR with a fluorescent tag

View ORCID ProfileLeigh A Stoddart, Nicholas D Kindon, Omolade Otun, Clare R. Harwood, Foteini Patera, View ORCID ProfileDmitry B Veprintsev, View ORCID ProfileJeanette Woolard, View ORCID ProfileStephen J Briddon, View ORCID ProfileHester A Franks, View ORCID ProfileStephen J Hill, View ORCID ProfileBarrie Kellam
doi: https://doi.org/10.1101/2020.04.21.053405
Leigh A Stoddart
1Cell Signalling and Pharmacology Research Group, Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham, UK
2Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Midlands, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Leigh A Stoddart
Nicholas D Kindon
2Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Midlands, UK
3School of Pharmacy, Biodiscovery Institute, University of Nottingham, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Omolade Otun
1Cell Signalling and Pharmacology Research Group, Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham, UK
2Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Midlands, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Clare R. Harwood
1Cell Signalling and Pharmacology Research Group, Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham, UK
2Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Midlands, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Foteini Patera
4Division of Cancer and Stem Cells, School of Medicine, University of Nottingham, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Dmitry B Veprintsev
1Cell Signalling and Pharmacology Research Group, Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham, UK
2Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Midlands, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Dmitry B Veprintsev
Jeanette Woolard
1Cell Signalling and Pharmacology Research Group, Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham, UK
2Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Midlands, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Jeanette Woolard
Stephen J Briddon
1Cell Signalling and Pharmacology Research Group, Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham, UK
2Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Midlands, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Stephen J Briddon
Hester A Franks
2Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Midlands, UK
4Division of Cancer and Stem Cells, School of Medicine, University of Nottingham, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Hester A Franks
Stephen J Hill
1Cell Signalling and Pharmacology Research Group, Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham, UK
2Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Midlands, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Stephen J Hill
  • For correspondence: stephen.hill@nottingham.ac.uk barrie.kellam@nottingham.ac.uk
Barrie Kellam
2Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Midlands, UK
3School of Pharmacy, Biodiscovery Institute, University of Nottingham, UK
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Barrie Kellam
  • For correspondence: stephen.hill@nottingham.ac.uk barrie.kellam@nottingham.ac.uk
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Supplementary material
  • Preview PDF
Loading

Abstract

Here, we describe rational design of a compound that covalently and selectively labels a G protein-coupled receptor (GPCR) in living cells with a fluorescent moiety. Using wild-type adenosine A2A receptor as a model system, we show fluorescent labelling without impeding access to the orthosteric binding site and demonstrate its use in endogenously expressing systems. This offers a non-invasive and selective approach to study function and localisation of native GPCRs.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
Back to top
PreviousNext
Posted April 23, 2020.
Download PDF

Supplementary Material

Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
Ligand-directed covalent labelling of a GPCR with a fluorescent tag
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Ligand-directed covalent labelling of a GPCR with a fluorescent tag
Leigh A Stoddart, Nicholas D Kindon, Omolade Otun, Clare R. Harwood, Foteini Patera, Dmitry B Veprintsev, Jeanette Woolard, Stephen J Briddon, Hester A Franks, Stephen J Hill, Barrie Kellam
bioRxiv 2020.04.21.053405; doi: https://doi.org/10.1101/2020.04.21.053405
Reddit logo Twitter logo Facebook logo LinkedIn logo Mendeley logo
Citation Tools
Ligand-directed covalent labelling of a GPCR with a fluorescent tag
Leigh A Stoddart, Nicholas D Kindon, Omolade Otun, Clare R. Harwood, Foteini Patera, Dmitry B Veprintsev, Jeanette Woolard, Stephen J Briddon, Hester A Franks, Stephen J Hill, Barrie Kellam
bioRxiv 2020.04.21.053405; doi: https://doi.org/10.1101/2020.04.21.053405

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Pharmacology and Toxicology
Subject Areas
All Articles
  • Animal Behavior and Cognition (4658)
  • Biochemistry (10313)
  • Bioengineering (7636)
  • Bioinformatics (26241)
  • Biophysics (13481)
  • Cancer Biology (10650)
  • Cell Biology (15361)
  • Clinical Trials (138)
  • Developmental Biology (8464)
  • Ecology (12776)
  • Epidemiology (2067)
  • Evolutionary Biology (16794)
  • Genetics (11373)
  • Genomics (15431)
  • Immunology (10580)
  • Microbiology (25087)
  • Molecular Biology (10172)
  • Neuroscience (54233)
  • Paleontology (398)
  • Pathology (1660)
  • Pharmacology and Toxicology (2884)
  • Physiology (4326)
  • Plant Biology (9213)
  • Scientific Communication and Education (1582)
  • Synthetic Biology (2545)
  • Systems Biology (6761)
  • Zoology (1459)