Modular design of cys-loop ligand-gated ion channels: Functional 5-HT 3 and GABA ρ1 receptors lacking the large cytoplasmic M3M4 loop

Michaela Jansen, Moez Bali, Myles Akabas

Research output: Contribution to journalArticle

74 Citations (Scopus)

Abstract

Cys-loop receptor neurotransmitter-gated ion channels are pentameric assemblies of subunits that contain three domains: extracellular, transmembrane, and intracellular. The extracellular domain forms the agonist binding site. The transmembrane domain forms the ion channel. The cytoplasmic domain is involved in trafficking, localization, and modulation by cytoplasmic second messenger systems but its role in channel assembly and function is poorly understood and little is known about its structure. The intracellular domain is formed by the large (> 100 residues) loop between the α-helical M3 and M4 transmembrane segments. Putative prokaryotic Cys-loop homologues lack a large M3M4 loop. We replaced the complete M3M4 loop (115 amino acids) in the 5-hydroxytryptamine type 3A (5-HT3A) subunit with a heptapeptide from the prokaryotic homologue from Gloeobacter violaceus. The macroscopic electrophysiological and pharmacological characteristics of the homomeric 5-HT3A-glvM3M4 receptors were comparable to 5-HT3A wild type. The channels remained cation-selective but the 5-HT3A-glvM3M4 single channel conductance was 43.5 pS as compared with the subpicosiemens wild-type conductance. Coexpression of hRIC-3, a protein that modulates expression of 5-HT3 and acetylcholine receptors, significantly attenuated 5-HT-induced currents with wild-type 5-HT3A but not 5-HT3A-glvM3M4 receptors. A similar deletion of the M3M4 loop in the anion-selective GABA-ρ1 receptor yielded functional, GABA-activated, anion-selective channels. These results imply that the M3M4 loop is not essential for receptor assembly and function and suggest that the cytoplasmic domain may fold as an independent module from the transmembrane and extracellular domains.

Original languageEnglish (US)
Pages (from-to)137-146
Number of pages10
JournalJournal of General Physiology
Volume131
Issue number2
DOIs
StatePublished - Feb 2008

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Ligand-Gated Ion Channels
GABA Receptors
Serotonin
Ion Channels
Anions
Cysteine Loop Ligand-Gated Ion Channel Receptors
Receptors, Serotonin, 5-HT3
Second Messenger Systems
Cholinergic Receptors
gamma-Aminobutyric Acid
Neurotransmitter Agents
Cations
Binding Sites
Pharmacology
Amino Acids

ASJC Scopus subject areas

  • Physiology

Cite this

Modular design of cys-loop ligand-gated ion channels : Functional 5-HT 3 and GABA ρ1 receptors lacking the large cytoplasmic M3M4 loop. / Jansen, Michaela; Bali, Moez; Akabas, Myles.

In: Journal of General Physiology, Vol. 131, No. 2, 02.2008, p. 137-146.

Research output: Contribution to journalArticle

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