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J Neurochem
2016 May 01;1374:528-38. doi: 10.1111/jnc.13578.
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Direct interaction of the resistance to inhibitors of cholinesterase type 3 protein with the serotonin receptor type 3A intracellular domain.
Nishtala SN
,
Mnatsakanyan N
,
Pandhare A
,
Leung C
,
Jansen M
.
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Pentameric ligand-gated ion channels (pLGIC) are expressed in both excitable and non-excitable cells that are targeted by numerous clinically used drugs. Assembly from five identical or homologous subunits yields homo- or heteromeric pentamers, respectively. The protein known as Resistance to Inhibitors of Cholinesterase (RIC-3) was identified to interfere with assembly and functional maturation of pLGICs. We have shown previously for serotonin type 3A homopentamers (5-HT3A ) that the interaction with RIC-3 requires the intracellular domain (ICD) of this pLGIC. After expression in Xenopus laevis oocytes RIC-3 attenuated serotonin-induced currents in 5-HT3A wild-type channels, but not in functional 5-HT3A glvM3M4 channels that have the 115-amino acid ICD replaced by a heptapeptide. In complementary experiments we have shown that engineering the Gloeobacter violaceus ligand-gated ion channel (GLIC) to contain the 5-HT3A -ICD confers sensitivity to RIC-3 in oocytes to otherwise insensitive GLIC. In this study, we identify endogenous RIC-3 protein expression in X. laevis oocytes. We purified RIC-3 to homogeneity after expression in Echericia coli. By using heterologously over-expressed and purified RIC-3 and the chimera consisting of the 5-HT3A -ICD and the extracellular and transmembrane domains of GLIC in pull-down experiments, we demonstrate a direct and specific interaction between the two proteins. This result further underlines that the domain within 5-HT3 A R that mediates the interaction with RIC-3 is the ICD. Importantly, this is the first experimental evidence that the interaction between 5-HT3 A R-ICD and RIC-3 does not require other proteins. In addition, we demonstrate that the pentameric assembly of the GLIC-5-HT3A -ICD chimera interacts with RIC-3. We hypothesized that pentameric ligand-gated ion channels (pLGICs) associate directly with the chaperone protein RIC-3 (resistance to inhibitors of cholinesterase type 3), and that the interaction does not require other protein factors. We found that the two proteins indeed interact directly, that the pLGIC intracellular domain is required for the effect, and that pLGICs in their pentameric form associate with RIC-3. These results provide the basis for future studies aimed at investigating which motifs provide the interaction surfaces, and at delineating the mechanism(s) of RIC-3 modulation of functional pLGIC surface expression.
Alexander,
Ric-3 promotes alpha7 nicotinic receptor assembly and trafficking through the ER subcompartment of dendrites.
2010, Pubmed
Alexander,
Ric-3 promotes alpha7 nicotinic receptor assembly and trafficking through the ER subcompartment of dendrites.
2010,
Pubmed
Baptista-Hon,
The minimum M3-M4 loop length of neurotransmitter-activated pentameric receptors is critical for the structural integrity of cytoplasmic portals.
2013,
Pubmed
Bar-Lev,
Molecular dissection of Cl--selective Cys-loop receptor points to components that are dispensable or essential for channel activity.
2011,
Pubmed
Bennett,
Xenopus laevis RIC-3 enhances the functional expression of the C. elegans homomeric nicotinic receptor, ACR-16, in Xenopus oocytes.
2012,
Pubmed
,
Xenbase
Biaggi-Labiosa,
Engineering α4β2 nAChRs with reduced or increased nicotine sensitivity via selective disruption of consensus sites in the M3-M4 cytoplasmic loop of the α4 subunit.
2015,
Pubmed
,
Xenbase
Blount,
BIP associates with newly synthesized subunits of the mouse muscle nicotinic receptor.
1991,
Pubmed
Bocquet,
A prokaryotic proton-gated ion channel from the nicotinic acetylcholine receptor family.
2007,
Pubmed
,
Xenbase
Bocquet,
X-ray structure of a pentameric ligand-gated ion channel in an apparently open conformation.
2009,
Pubmed
Castelán,
Molecular characterization and localization of the RIC-3 protein, an effector of nicotinic acetylcholine receptor expression.
2008,
Pubmed
,
Xenbase
Castillo,
Dual role of the RIC-3 protein in trafficking of serotonin and nicotinic acetylcholine receptors.
2005,
Pubmed
,
Xenbase
Cheng,
Cell surface expression of 5-hydroxytryptamine type 3 receptors is promoted by RIC-3.
2005,
Pubmed
Cheng,
Differential subcellular localization of RIC-3 isoforms and their role in determining 5-HT3 receptor composition.
2007,
Pubmed
Du,
Glycine receptor mechanism elucidated by electron cryo-microscopy.
2015,
Pubmed
Gelman,
Role of the endoplasmic reticulum chaperone calnexin in subunit folding and assembly of nicotinic acetylcholine receptors.
1995,
Pubmed
Gonzalez-Gutierrez,
Mutations that stabilize the open state of the Erwinia chrisanthemi ligand-gated ion channel fail to change the conformation of the pore domain in crystals.
2012,
Pubmed
,
Xenbase
Goyal,
Engineering a prokaryotic Cys-loop receptor with a third functional domain.
2011,
Pubmed
,
Xenbase
Green,
Ion channel assembly: creating structures that function.
1999,
Pubmed
Halevi,
The C. elegans ric-3 gene is required for maturation of nicotinic acetylcholine receptors.
2002,
Pubmed
,
Xenbase
Halevi,
Conservation within the RIC-3 gene family. Effectors of mammalian nicotinic acetylcholine receptor expression.
2003,
Pubmed
,
Xenbase
Hassaine,
X-ray structure of the mouse serotonin 5-HT3 receptor.
2014,
Pubmed
Hilf,
X-ray structure of a prokaryotic pentameric ligand-gated ion channel.
2008,
Pubmed
Hilf,
Structure of a potentially open state of a proton-activated pentameric ligand-gated ion channel.
2009,
Pubmed
Huebsch,
Rapsyn-mediated clustering of acetylcholine receptor subunits requires the major cytoplasmic loop of the receptor subunits.
2003,
Pubmed
Jansen,
Modular design of Cys-loop ligand-gated ion channels: functional 5-HT3 and GABA rho1 receptors lacking the large cytoplasmic M3M4 loop.
2008,
Pubmed
,
Xenbase
Kabbani,
Are nicotinic acetylcholine receptors coupled to G proteins?
2013,
Pubmed
Langlhofer,
Length of the TM3-4 loop of the glycine receptor modulates receptor desensitization.
2015,
Pubmed
Lansdell,
RIC-3 enhances functional expression of multiple nicotinic acetylcholine receptor subtypes in mammalian cells.
2005,
Pubmed
,
Xenbase
Lansdell,
Host-cell specific effects of the nicotinic acetylcholine receptor chaperone RIC-3 revealed by a comparison of human and Drosophila RIC-3 homologues.
2008,
Pubmed
McKinnon,
5-HT3 receptor ion size selectivity is a property of the transmembrane channel, not the cytoplasmic vestibule portals.
2011,
Pubmed
,
Xenbase
McKinnon,
Length and amino acid sequence of peptides substituted for the 5-HT3A receptor M3M4 loop may affect channel expression and desensitization.
2012,
Pubmed
,
Xenbase
Melzer,
Multifunctional basic motif in the glycine receptor intracellular domain induces subunit-specific sorting.
2010,
Pubmed
Meyer,
Identification of a gephyrin binding motif on the glycine receptor beta subunit.
1995,
Pubmed
Millar,
Assembly and trafficking of nicotinic acetylcholine receptors (Review).
2008,
Pubmed
Miller,
Crystal structure of a human GABAA receptor.
2014,
Pubmed
Mnatsakanyan,
Functional Chimeras of GLIC Obtained by Adding the Intracellular Domain of Anion- and Cation-Conducting Cys-Loop Receptors.
2015,
Pubmed
Moraga-Cid,
A Single phenylalanine residue in the main intracellular loop of α1 γ-aminobutyric acid type A and glycine receptors influences their sensitivity to propofol.
2011,
Pubmed
Moroni,
Chloride ions in the pore of glycine and GABA channels shape the time course and voltage dependence of agonist currents.
2011,
Pubmed
Pan,
Structure of the pentameric ligand-gated ion channel ELIC cocrystallized with its competitive antagonist acetylcholine.
2012,
Pubmed
Pan,
Structure of the pentameric ligand-gated ion channel GLIC bound with anesthetic ketamine.
2012,
Pubmed
,
Xenbase
Papke,
The role of intracellular linkers in gating and desensitization of human pentameric ligand-gated ion channels.
2014,
Pubmed
Ramarao,
Mechanism of nicotinic acetylcholine receptor cluster formation by rapsyn.
1998,
Pubmed
Recchia,
Alpha-synuclein and Parkinson's disease.
2004,
Pubmed
Riggs,
Expression and purification of recombinant proteins by fusion to maltose-binding protein.
2000,
Pubmed
Roncarati,
Functional properties of alpha7 nicotinic acetylcholine receptors co-expressed with RIC-3 in a stable recombinant CHO-K1 cell line.
2008,
Pubmed
Seddon,
Membrane proteins, lipids and detergents: not just a soap opera.
2004,
Pubmed
Swope,
Regulation of ligand-gated ion channels by protein phosphorylation.
1999,
Pubmed
Tasneem,
Identification of the prokaryotic ligand-gated ion channels and their implications for the mechanisms and origins of animal Cys-loop ion channels.
2005,
Pubmed
Tretter,
The clustering of GABA(A) receptor subtypes at inhibitory synapses is facilitated via the direct binding of receptor alpha 2 subunits to gephyrin.
2008,
Pubmed
Unterer,
The importance of TM3-4 loop subdomains for functional reconstitution of glycine receptors by independent domains.
2012,
Pubmed
Vallés,
Ric-3 chaperone-mediated stable cell-surface expression of the neuronal alpha7 nicotinic acetylcholine receptor in mammalian cells.
2009,
Pubmed
Vallés,
Chaperoning α7 neuronal nicotinic acetylcholine receptors.
2012,
Pubmed
Vithlani,
The dynamic modulation of GABA(A) receptor trafficking and its role in regulating the plasticity of inhibitory synapses.
2011,
Pubmed
Williams,
Ric-3 promotes functional expression of the nicotinic acetylcholine receptor alpha7 subunit in mammalian cells.
2005,
Pubmed
,
Xenbase
Yevenes,
Modulation of glycine-activated ion channel function by G-protein betagamma subunits.
2003,
Pubmed
Yu,
A sequence in the main cytoplasmic loop of the alpha subunit is required for assembly of mouse muscle nicotinic acetylcholine receptor.
1994,
Pubmed