Development of novel fluorescent histamine H₁-receptor antagonists to study ligand-binding kinetics in living cells

The histamine H1-receptor (H1R) is an important mediator of allergy and inflammation. H1R antagonists have particular clinical utility in allergic rhinitis and urticaria. Here we have developed six novel fluorescent probes for this receptor that are very effective for high resolution confocal imagin...

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Main Authors: Stoddart, Leigh A., Vernall, Andrea J., Bouzo-Lorenzo, Monica, Bosma, Reggie, Kooistra, Albert J., de Graaf, Chris, Vischer, Henry F., Leurs, Rob, Briddon, Stephen J., Kellam, Barrie, Hill, Stephen J.
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Published: Nature Publishing Group 2018
Online Access:https://eprints.nottingham.ac.uk/49204/
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author Stoddart, Leigh A.
Vernall, Andrea J.
Bouzo-Lorenzo, Monica
Bosma, Reggie
Kooistra, Albert J.
de Graaf, Chris
Vischer, Henry F.
Leurs, Rob
Briddon, Stephen J.
Kellam, Barrie
Hill, Stephen J.
author_facet Stoddart, Leigh A.
Vernall, Andrea J.
Bouzo-Lorenzo, Monica
Bosma, Reggie
Kooistra, Albert J.
de Graaf, Chris
Vischer, Henry F.
Leurs, Rob
Briddon, Stephen J.
Kellam, Barrie
Hill, Stephen J.
author_sort Stoddart, Leigh A.
building Nottingham Research Data Repository
collection Online Access
description The histamine H1-receptor (H1R) is an important mediator of allergy and inflammation. H1R antagonists have particular clinical utility in allergic rhinitis and urticaria. Here we have developed six novel fluorescent probes for this receptor that are very effective for high resolution confocal imaging, alongside bioluminescence resonance energy transfer approaches to monitor H1R ligand binding kinetics in living cells. The latter technology exploits the opportunities provided by the recently described bright bioluminescent protein NanoLuc when it is fused to the N-terminus of a receptor. Two different pharmacophores (mepyramine or the fragment VUF13816) were used to generate fluorescent H1R antagonists conjugated via peptide linkers to the fluorophore BODIPY630/650. Kinetic properties of the probes showed wide variation, with the VUF13816 analogues having much longer H1R residence times relative to their mepyramine-based counterparts. The kinetics of these fluorescent ligands could also be monitored in membrane preparations providing new opportunities for future drug discovery applications.
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publishDate 2018
publisher Nature Publishing Group
recordtype eprints
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spelling nottingham-492042020-05-04T19:28:03Z https://eprints.nottingham.ac.uk/49204/ Development of novel fluorescent histamine H₁-receptor antagonists to study ligand-binding kinetics in living cells Stoddart, Leigh A. Vernall, Andrea J. Bouzo-Lorenzo, Monica Bosma, Reggie Kooistra, Albert J. de Graaf, Chris Vischer, Henry F. Leurs, Rob Briddon, Stephen J. Kellam, Barrie Hill, Stephen J. The histamine H1-receptor (H1R) is an important mediator of allergy and inflammation. H1R antagonists have particular clinical utility in allergic rhinitis and urticaria. Here we have developed six novel fluorescent probes for this receptor that are very effective for high resolution confocal imaging, alongside bioluminescence resonance energy transfer approaches to monitor H1R ligand binding kinetics in living cells. The latter technology exploits the opportunities provided by the recently described bright bioluminescent protein NanoLuc when it is fused to the N-terminus of a receptor. Two different pharmacophores (mepyramine or the fragment VUF13816) were used to generate fluorescent H1R antagonists conjugated via peptide linkers to the fluorophore BODIPY630/650. Kinetic properties of the probes showed wide variation, with the VUF13816 analogues having much longer H1R residence times relative to their mepyramine-based counterparts. The kinetics of these fluorescent ligands could also be monitored in membrane preparations providing new opportunities for future drug discovery applications. Nature Publishing Group 2018-01-25 Article PeerReviewed Stoddart, Leigh A., Vernall, Andrea J., Bouzo-Lorenzo, Monica, Bosma, Reggie, Kooistra, Albert J., de Graaf, Chris, Vischer, Henry F., Leurs, Rob, Briddon, Stephen J., Kellam, Barrie and Hill, Stephen J. (2018) Development of novel fluorescent histamine H₁-receptor antagonists to study ligand-binding kinetics in living cells. Scientific Reports, 8 . p. 1572. ISSN 2045-2322 https://www.nature.com/articles/s41598-018-19714-2 doi:10.1038/s41598-018-19714-2 doi:10.1038/s41598-018-19714-2
spellingShingle Stoddart, Leigh A.
Vernall, Andrea J.
Bouzo-Lorenzo, Monica
Bosma, Reggie
Kooistra, Albert J.
de Graaf, Chris
Vischer, Henry F.
Leurs, Rob
Briddon, Stephen J.
Kellam, Barrie
Hill, Stephen J.
Development of novel fluorescent histamine H₁-receptor antagonists to study ligand-binding kinetics in living cells
title Development of novel fluorescent histamine H₁-receptor antagonists to study ligand-binding kinetics in living cells
title_full Development of novel fluorescent histamine H₁-receptor antagonists to study ligand-binding kinetics in living cells
title_fullStr Development of novel fluorescent histamine H₁-receptor antagonists to study ligand-binding kinetics in living cells
title_full_unstemmed Development of novel fluorescent histamine H₁-receptor antagonists to study ligand-binding kinetics in living cells
title_short Development of novel fluorescent histamine H₁-receptor antagonists to study ligand-binding kinetics in living cells
title_sort development of novel fluorescent histamine h₁-receptor antagonists to study ligand-binding kinetics in living cells
url https://eprints.nottingham.ac.uk/49204/
https://eprints.nottingham.ac.uk/49204/
https://eprints.nottingham.ac.uk/49204/