An allosteric binding site of the α7 nicotinic acetylcholine receptor revealed in a humanized acetylcholine-binding protein

• DOI: 10.1074/jbc.m117.815316
• Publication Date: 2017-12-13T17:15:19Z
• Authors: Florian Delbart; Marijke Brams; Fabian Gruss; Sam Noppen; Steve Peigneur; Sandro Boland; Patrick Chaltin; Jose Brandao-Neto; Frank von Delft; Wouter G. Touw; Robbie P. Joosten; Sandra Liekens; Jan Tytgat; Chris Ulens
• Keywords: Models, Molecular; 0301 basic medicine; alpha7 Nicotinic Acetylcholine Receptor; Snails; Receptors, Nicotinic; Crystallography, X-Ray; Acetylcholine; 3. Good health; 03 medical and health sciences; Neurobiology; Allosteric Regulation; Protein Domains; Animals; Humans; Allosteric Site

Nicotinic acetylcholine receptors (nAChRs) belong to the family of pentameric ligand-gated ion channels and mediate fast excitatory transmission in the central and peripheral nervous systems. Among the different existing receptor subtypes, the homomeric α7 nAChR has attracted considerable attention because of its possible implication in several neurological and psychiatric disorders, including cognitive decline associated with Alzheimer's disease or schizophrenia. Allosteric modulators of ligand-gated ion channels are of particular interest as therapeutic agents, as they modulate receptor activity without affecting normal fluctuations of synaptic neurotransmitter release. Here, we used X-ray crystallography and surface plasmon resonance spectroscopy of α7-acetylcholine-binding protein (AChBP), a humanized chimera of a snail AChBP, which has 71% sequence similarity with the extracellular ligand-binding domain of the human α7 nAChR, to investigate the structural determinants of allosteric modulation. We extended previous observations that an allosteric site located in the vestibule of the receptor offers an attractive target for receptor modulation. We introduced seven additional humanizing mutations in the vestibule-located binding site of AChBP to improve its suitability as a model for studying allosteric binding. Using a fragment-based screening approach, we uncovered an allosteric binding site located near the β8-β9 loop, which critically contributes to coupling ligand binding to channel opening in human α7 nAChR. This work expands our understanding of the topology of allosteric binding sites in AChBP and, by extrapolation, in the human α7 nAChR as determined by electrophysiology measurements. Our insights pave the way for drug design strategies targeting nAChRs involved in ion channel-mediated disorders.