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MOP and NOP receptor interaction: Studies with a dual expression system and bivalent peptide ligands

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posted on 2022-05-20, 10:26 authored by Mark F Bird, J. McDonald, B. Horley, J. P. O’Doherty, B. Fraser, C. L. Gibson, R. Guerrini, David Lambert

Opioids targeting mu;μ (MOP) receptors produce analgesia in the peri-operative period and palliative care. They also produce side effects including respiratory depression, tolerance/dependence and addiction. The N/OFQ opioid receptor (NOP) also produces analgesia but is devoid of the major MOP side effects. Evidence exists for MOP-NOP interaction and mixed MOP-NOP ligands produce analgesia with reduced side effects. We have generated a HEKMOP/NOP human expression system and used bivalent MOP-NOP and fluorescent ligands to (i) probe for receptor interaction and (ii) consequences of that interaction. We used HEKMOP/NOP cells and two bivalent ligands; Dermorphin-N/OFQ (MOP agonist-NOP agonist; DeNO) and Dermorphin-UFP101 (MOP agonist-NOP antagonist; De101). We have determined receptor binding profiles, GTPγ[35S] binding, cAMP formation and ERK1/2 activation. We have also probed MOP and NOP receptor interactions in HEK cells and hippocampal neurones using the novel MOP fluorescent ligand, DermorphinATTO488 and the NOP fluorescent ligand N/OFQATTO594. In HEKMOP/NOP MOP ligands displaced NOP binding and NOP ligands displaced MOP binding. Using fluorescent probes in HEKMOP/NOP cells we demonstrated MOP-NOP probe overlap and a FRET signal indicating co-localisation. MOP-NOP were also co-localised in hippocampal tissue. In GTPγ[35S] and cAMP assays NOP stimulation shifted the response to MOP rightwards. At ERK1/2 the response to bivalent ligands generally peaked later. We provide evidence for MOP-NOP interaction in recombinant and native tissue. NOP activation reduces responsiveness of MOP activation; this was shown with conventional and bivalent ligands. 

Funding

British Journal of Anaesthesia

History

Citation

PLoS ONE 17(1): e0260880. https://doi.org/10.1371/journal.pone.0260880

Author affiliation

Department of Cardiovascular Sciences, University of Leicester

Version

  • VoR (Version of Record)

Published in

PLoS One

Volume

17

Issue

1

Publisher

Public Library of Science (PLoS)

issn

1932-6203

Acceptance date

2021-11-18

Copyright date

2022

Available date

2022-05-20

Language

en

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