Generation of nanobodies acting as silent and positive allosteric modulators of the α7 nicotinic acetylcholine receptor
| Autor: | Qimeng Li, Ákos Nemecz, Gabriel Aymé, Gabrielle Dejean de la Bâtie, Marie S. Prevost, Stéphanie Pons, Nathalie Barilone, Rayen Baachaoui, Uwe Maskos, Pierre Lafaye, Pierre-Jean Corringer |
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| Přispěvatelé: | Récepteurs Canaux - Channel Receptors, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Lanzhou Institute of Biological Products Co. Ltd. (LIBP), Collège Doctoral, Sorbonne Université (SU), Ingénierie des Anticorps (plate-forme) - Antibody Engineering (Platform), Neurobiologie intégrative des Systèmes cholinergiques / Integrative Neurobiology of Cholinergic Systems (NISC), Institut Pasteur [Paris] (IP)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), grant from LanZhou Institute of Biological Products Co., Ltd, LanZhou, China, Institut Pasteur PPU program, Institut Pasteur 'Programme transversal de recherche' (PTR 03-17 Nicobinder), Institut National du Cancer' INCa, Fondation pour la Recherche Médicale' FRM-Equipe, INCa, ERA-NET Neuron, Centre National de la Recherche Scientifique, doctoral school ED3C, Sylvie Bay for help in nanobody characterization by mass spectroscopy, Pierre-Henri Commere for help in flow-cytometry experiments |
| Jazyk: | angličtina |
| Rok vydání: | 2023 |
| Předmět: |
Pharmacology
Cellular and Molecular Neuroscience nanobody α7-nAChR nicotinic acetylcholine receptor VHH nanobody allosteric modulator [SDV]Life Sciences [q-bio] Molecular Medicine allosteric modulator [CHIM]Chemical Sciences α7-nAChR VHH Cell Biology nicotinic acetylcholine receptor Molecular Biology |
| Popis: | The α7 nicotinic acetylcholine receptor (nAChR), a potential drug target for treating cognitive disorders, mediates communication between neuronal and non-neuronal cells. Although many competitive antagonists, agonists, and partial-agonists have been found and synthesized, they have not led to effective therapeutic treatments. In this context, small molecules acting as positive allosteric modulators binding outside the orthosteric, acetylcholine, site have attracted considerable interest. Two single-domain antibody fragments, C4 and E3, against the extracellular domain of the human α7-nAChR were generated through alpaca immunization with cells expressing a human α7-nAChR/mouse 5-HT3A chimera, and are herein described. They bind to the α7-nAChR but not to the other major nAChR subtypes, α4β2 and α3β4. E3 acts as a slowly associating positive allosteric modulator, strongly potentiating the acetylcholine-elicited currents, while not precluding the desensitization of the receptor. An E3–E3 bivalent construct shows similar potentiating properties but displays very slow dissociation kinetics conferring quasi-irreversible properties. Whereas, C4 does not alter the receptor function, but fully inhibits the E3-evoked potentiation, showing it is a silent allosteric modulator competing with E3 binding. Both nanobodies do not compete with α-bungarotoxin, localizing at an allosteric extracellular binding site away from the orthosteric site. The functional differences of each nanobody, as well as the alteration of functional properties through nanobody modifications indicate the importance of this extracellular site. The nanobodies will be useful for pharmacological and structural investigations; moreover, they, along with the extracellular site, have a direct potential for clinical applications. |
| Databáze: | OpenAIRE |
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