Structural Basis of μ-Opioid Receptor-Targeting by a Nanobody Antagonist

Article Article scientifique Article Structural Basis of μ-Opioid Receptor-Targeting by a Nanobody Antagonist eng 2023-12-07 2023 author Yu Jun 924631 Département de biologie moléculaire et cellulaire (MOCEL) author Kumar Amit 924488 Département de physiologie cellulaire et métabolisme (PHYME) author Zhang Xuefeng 0000-0002-5222-7042 402287 Département de biologie moléculaire et cellulaire (MOCEL) author Martin Charlotte 842564 author Raia Pierre 930734 Département des sciences végétales (BIVEG) author Koehl Antoine author Laeremans Toon author Steyaert Jan author Manglik Aashish author Ballet Steven author Boland Andreas 912539 Département de biologie moléculaire et cellulaire (MOCEL) author Stoeber Miriam Carolin 930635 Département de physiologie cellulaire et métabolisme (PHYME) 10.1101/2023.12.06.570395 38106026 1559-6095 PMC10723425 http://biorxiv.org/lookup/doi/10.1101/2023.12.06.570395 <p>The μ-opioid receptor (μOR), a prototypical member of the G protein-coupled receptor (GPCR) family, is the molecular target of opioid analgesics such as morphine and fentanyl. Due to the limitations and severe side effects of currently available opioid drugs, there is considerable interest in developing novel modulators of μOR function. Most GPCR ligands today are small molecules, however biologics, including antibodies and nanobodies, are emerging as alternative therapeutics with clear advantages such as affinity and target selectivity. Here, we describe the nanobody NbE, which selectively binds to the μOR and acts as an antagonist. We functionally characterize NbE as an extracellular and genetically encoded µOR ligand and uncover the molecular basis for µOR antagonism by solving the cryo-EM structure of the NbE-µOR complex. NbE displays a unique ligand binding mode and achieves µOR selectivity by interactions with the orthosteric pocket and extracellular receptor loops. Based on a β-hairpin loop formed by NbE that deeply inserts into the µOR and centers most binding contacts, we design short peptide analogues that retain µOR antagonism. The work illustrates the potential of nanobodies to uniquely engage with GPCRs and describes novel μOR ligands that can serve as a basis for therapeutic developments.</p> Dewey 580 Dewey 570 45 Département des sciences végétales 163 7daed2ee-1311-4824-9df9-2e51260a966a Département de biologie moléculaire et cellulaire 16484 false Cold Spring Harbor protocols 97d4be0c-b047-47dd-8cb4-d75b1f6b77cc Article (Submitted version) Public BOLAND01.pdf 12038117 application/pdf CC BY-NC-ND-4.0 27d66a15ce103f0a57e3e0c941fe5c17ec2e1100f278d954c1f20a8ba3886d44 openAccess Faculté des sciences / Section de biologie / Département des sciences végétales Faculté des sciences / Section de biologie / Département de biologie moléculaire et cellulaire

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Structural Basis of μ-Opioid Receptor-Targeting by a Nanobody Antagonist	Cold Spring Harbor protocols			2023	85	53

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