The role of intracellular linkers in gating and desensitization of human pentameric ligand-gated ion channels

David Papke, Claudio Grosman

Research output: Contribution to journalArticle

Abstract

It has recently been proposed that post-translational modification of not only the M3-M4 linker but also the M1-M2 linker of pentameric ligand-gated ion channels modulates function in vivo. To estimate the involvement of the M1-M2 linker in gating and desensitization, we engineered a series of mutations to this linker of the human adult-muscle acetylcholine receptor (AChR), the γ3 4 AChR and the homomeric γ1 glycine receptor (GlyR). All tested M1-M2 linker mutations had little effect on the kinetics of deactivation or desensitization compared with the effects of mutations to the M2 γ-helix or the extracellular M2-M3 linker. However, when the effects of mutations were assessed with 50 Hz trains of 1 ms pulses of saturating neurotransmitter, some mutations led to much more, and others to much less, peak-current depression than observed for the wild-type channels, suggesting that these mutations could affect the fidelity of fast synaptic transmission. Nevertheless, no mutation to this linker could mimic the irreversible loss of responsiveness reported to result from the oxidation of the M1-M2 linker cysteines of the γ3 AChR subunit. We also replaced the M3-M4 linker of the γ1 GlyR with much shorter peptides and found that none of these extensive changes affects channel deactivation strongly or reduces the marked variability in desensitization kinetics that characterizes the wild-type channel. However, we found that these large mutations to the M3-M4 linker can have pronounced effects on desensitization kinetics, supporting the notion that its post-translational modification could indeed modulate γ1 GlyR behavior.

Original languageEnglish (US)
Pages (from-to)7238-7252
Number of pages15
JournalJournal of Neuroscience
Volume34
Issue number21
DOIs
StatePublished - Jan 1 2014

Keywords

  • Fast perfusion
  • Glycine receptors
  • Ion-channel kinetics
  • Nicotinic receptors
  • Outside-out patches
  • Patch-clamp

ASJC Scopus subject areas

  • Neuroscience(all)

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