title: Incompatibility between a pair of residues from the pre-M1 linker and Cys-loop blocks surface expression of the glycine receptor.
abstract: Regulation of cell membrane excitability can be achieved either by modulating the functional properties of cell membrane-expressed single channels or by varying the number of expressed channels. Whereas the structural basis underlying single channel properties has been intensively studied, the structural basis contributing to surface expression is less well characterized. Here we demonstrate that homologous substitution of the pre-M1 linker from the β subunit prevents surface expression of the α1 glycine receptor chloride channel. By investigating a series of chimeras comprising α1 and β subunits, we hypothesized that this effect was due to incompatibility between a pair of positively charged residues, which lie in close proximity to each other in the tertiary structure, from the pre-M1 linker and Cys-loop. Abolishing either positive charge restored surface expression. We propose that incompatibility (electrostatic repulsion) between this pair of residues misfolds the glycine receptor, and in consequence, the protein is retained in the cytoplasm and prevented from surface expression by the quality control machinery. This hypothesis suggests a novel mechanism, i.e. residue incompatibility, for explaining the mutation-induced reduction in channel surface expression, often present in the cases of hereditary hyperekplexia.
authors.affiliation: Brain and Mind Research Institute, University of Sydney, Sydney, New South Wales, 2050 Australia. qshan@yahoo.com
authors.forename: Qiang
authors.initials: Q
authors.lastname: Shan
authors.forename: Joseph W
authors.initials: JW
authors.lastname: Lynch
mesh_terms: D019943:Amino Acid Substitution; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D020125:Mutation, Missense; D017433:Protein Structure, Secondary; D018009:Receptors, Glycine / Q000096:biosynthesis* / Q000235:genetics
chemical_list: C114561:GLRB protein, human; D018009:Receptors, Glycine