Structure of ligand-gated ion channels: critical assessment of biochemical data supports novel topology

Abstract

Rapid signaling across the synaptic junction is partially mediated by the ligand-gated ion channel superfamily (LGICS), which includes inhibitory glycine and GABA receptors and excitatory acetylcholine and serotonin receptors. The glycine receptor (GlyR) can assemble as homopentamers of alpha subunits, and baculovirus expression systems are capable of overexpressing large quantities of active receptors. Limited proteolysis coupled to mass spectrometry on reconstituted alpha1 GlyR homopentamers identified proteolytic cleavages within proposed transmembrane domains postulated to fold as bilayer-spanning alpha helices in the "classical" model and identified unexpected membrane-associated regions in the N-terminal domain (J. F. Leite et al., 2000, J. Biol. Chem. 275, 13683-13689). In this review, optimized sequence alignments were used to integrate these proteolysis data with biochemical information determined in studies of all the LGICS members in order to construct a novel topological model.