Asymmetric contribution of alpha and beta subunits to the activation of alphabeta heteromeric glycine receptors

• Published in: Journal of neurochemistry Vol. 86; no. 2; p. 498
• Main Authors: Shan, Qiang, Nevin, Simon T, Haddrill, Justine L, Lynch, Joseph W
• Format: Journal Article
• Language: English
• Published: England 01.07.2003
• Subjects: Binding, Competitive - drug effects; Binding, Competitive - physiology; Cell Line; Glycine - metabolism; Glycine - pharmacokinetics; Humans; Kidney - cytology; Kidney - metabolism; Mutagenesis, Site-Directed; Patch-Clamp Techniques; Protein Subunits - genetics; Protein Subunits - metabolism; Receptors, Glycine - genetics; Receptors, Glycine - metabolism; Structure-Activity Relationship; Transfection
• ISSN: 0022-3042

This study investigated the role of beta subunits in the activation of alphabeta heteromeric glycine receptor (GlyR) chloride channels recombinantly expressed in HEK293 cells. The approach involved incorporating mutations into corresponding positions in alpha and beta subunits and comparing their effects on receptor function. Although cysteine-substitution mutations to residues in the N-terminal half of the alpha subunit M2-M3 loop dramatically impaired the gating efficacy, the same mutations exerted little effect when incorporated into corresponding positions of the beta subunit. Furthermore, although the alpha subunit M2-M3 loop cysteines were modified by a cysteine-specific reagent, the corresponding beta subunit cysteines showed no evidence of reactivity. These observations suggest structural or functional differences between alpha and beta subunit M2-M3 loops. In addition, a threonine-->leucine mutation at the 9' position in the beta subunit M2 pore-lining domain dramatically increased the glycine sensitivity. By analogy with the effects of the same mutation in other ligand-gated ion channels, it was concluded that the mutation affected the GlyR activation mechanism. This supports the idea that the GlyR beta subunit is involved in receptor gating. In conclusion, this study demonstrates that beta subunits contribute to the activation of the GlyR, but that their involvement in this process is significantly different to that of the alpha subunit.