Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Insect Mol Biol
2022 Oct 01;315:620-633. doi: 10.1111/imb.12791.
Show Gene links
Show Anatomy links
Different efficiency of auxiliary/chaperone proteins to promote the functional reconstitution of honeybee glutamate and acetylcholine receptors in Xenopus laevis oocytes.
Brunello L
,
Ménard C
,
Rousset M
,
Vignes M
,
Charnet P
,
Cens T
.
???displayArticle.abstract???
Heterologous expression systems (e.g., Xenopus laevis oocytes) are useful to study the biophysical properties and pharmacology of ionotropic receptors such as ionotropic glutamate (iGLuRs) and nicotinic acetylcholine (nAChRs) receptors. However, insect receptors often require the co-expression of chaperone proteins to be functional. Only few iGluRs and nAChRs have been successfully expressed in such systems. Here, we compared the efficiency of chaperone proteins to promote the functional expression of one Apis mellifera iGluR and several nAChR subunit combinations (α1α8β1, α7, α2α8β1 and α2α7α8β1) in Xenopus oocytes. To this end, we cloned a new iGluR (GluR-1) and potential chaperone proteins (e.g., SOL-1, Neto, NACHO) and tested more than 40 combinations of human, nematode and honeybee proteins. We obtained robust expression of GluR-1 and α1α8β1 when co-expressed with honeybee chaperone proteins and found that nAChR expression critically depended on the α1 subunit N-terminal sequence. We recorded small ACh-gated currents in few oocytes when the α7 subunit was co-expressed with Caenorhabditis elegans RIC-3, but none of the chaperone proteins allowed efficient expression of α2α8β1 or α2α7α8β1. Our results show that only some protein combinations can reconstitute functional receptors in Xenopus oocytes and that protein combination efficient in one species is not always efficient in another species.
Ben-Ami,
RIC-3 affects properties and quantity of nicotinic acetylcholine receptors via a mechanism that does not require the coiled-coil domains.
2005, Pubmed,
Xenbase
Ben-Ami,
RIC-3 affects properties and quantity of nicotinic acetylcholine receptors via a mechanism that does not require the coiled-coil domains.
2005,
Pubmed
,
Xenbase
Ben-David,
RIC-3 expression and splicing regulate nAChR functional expression.
2016,
Pubmed
,
Xenbase
Bennett,
Xenopus laevis RIC-3 enhances the functional expression of the C. elegans homomeric nicotinic receptor, ACR-16, in Xenopus oocytes.
2012,
Pubmed
,
Xenbase
Biala,
The conserved RIC-3 coiled-coil domain mediates receptor-specific interactions with nicotinic acetylcholine receptors.
2009,
Pubmed
Boulin,
Eight genes are required for functional reconstitution of the Caenorhabditis elegans levamisole-sensitive acetylcholine receptor.
2008,
Pubmed
,
Xenbase
Brockie,
Ionotropic glutamate receptors in Caenorhabditis elegans.
2003,
Pubmed
Cartereau,
Cloning and Expression of Cockroach α7 Nicotinic Acetylcholine Receptor Subunit.
2020,
Pubmed
,
Xenbase
Castillo,
Dual role of the RIC-3 protein in trafficking of serotonin and nicotinic acetylcholine receptors.
2005,
Pubmed
,
Xenbase
Cens,
Modulation of the alpha 1A Ca2+ channel by beta subunits at physiological Ca2+ concentration.
1996,
Pubmed
,
Xenbase
Cens,
Molecular characterization and functional expression of the Apis mellifera voltage-dependent Ca2+ channels.
2015,
Pubmed
,
Xenbase
Chen,
Ecological toxicity reduction of dinotefuran to honeybee: New perspective from an enantiomeric level.
2019,
Pubmed
Dupuis,
Expression patterns of nicotinic subunits α2, α7, α8, and β1 affect the kinetics and pharmacology of ACh-induced currents in adult bee olfactory neuropiles.
2011,
Pubmed
Gally,
A transmembrane protein required for acetylcholine receptor clustering in Caenorhabditis elegans.
2004,
Pubmed
Gu,
Hair cell α9α10 nicotinic acetylcholine receptor functional expression regulated by ligand binding and deafness gene products.
2020,
Pubmed
Gu,
α6-Containing Nicotinic Acetylcholine Receptor Reconstitution Involves Mechanistically Distinct Accessory Components.
2019,
Pubmed
Gu,
Brain α7 Nicotinic Acetylcholine Receptor Assembly Requires NACHO.
2016,
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
Han,
Functional reconstitution of Drosophila melanogaster NMJ glutamate receptors.
2015,
Pubmed
,
Xenbase
Hansen,
Structure, Function, and Pharmacology of Glutamate Receptor Ion Channels.
2021,
Pubmed
Ihara,
Cofactor-enabled functional expression of fruit fly, honeybee, and bumblebee nicotinic receptors reveals picomolar neonicotinoid actions.
2020,
Pubmed
,
Xenbase
Jones,
InterProScan 5: genome-scale protein function classification.
2014,
Pubmed
Knowland,
Functional α6β4 acetylcholine receptor expression enables pharmacological testing of nicotinic agonists with analgesic properties.
2020,
Pubmed
Lansdell,
The Drosophila nicotinic acetylcholine receptor subunits Dα5 and Dα7 form functional homomeric and heteromeric ion channels.
2012,
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
Lu,
CDD/SPARCLE: the conserved domain database in 2020.
2020,
Pubmed
Nielsen,
A Brief History of Protein Sorting Prediction.
2019,
Pubmed
Rex,
A Genome-Wide Arrayed cDNA Screen to Identify Functional Modulators of α7 Nicotinic Acetylcholine Receptors.
2017,
Pubmed
Sandoval,
gamma1-dependent down-regulation of recombinant voltage-gated Ca2+ channels.
2007,
Pubmed
Seredenina,
Molecular cloning and characterization of a novel human variant of RIC-3, a putative chaperone of nicotinic acetylcholine receptors.
2008,
Pubmed
Shigetou,
Modulation by neonicotinoids of honeybee α1/chicken β2 hybrid nicotinic acetylcholine receptors expressed in Xenopus laevis oocytes.
2020,
Pubmed
Walker,
Reconstitution of invertebrate glutamate receptor function depends on stargazin-like proteins.
2006,
Pubmed
,
Xenbase
Walker,
Conserved SOL-1 proteins regulate ionotropic glutamate receptor desensitization.
2006,
Pubmed
Wang,
The SOL-2/Neto auxiliary protein modulates the function of AMPA-subtype ionotropic glutamate receptors.
2012,
Pubmed
,
Xenbase
Watson,
A spinosyn-sensitive Drosophila melanogaster nicotinic acetylcholine receptor identified through chemically induced target site resistance, resistance gene identification, and heterologous expression.
2010,
Pubmed
,
Xenbase
Williams,
Ric-3 promotes functional expression of the nicotinic acetylcholine receptor alpha7 subunit in mammalian cells.
2005,
Pubmed
,
Xenbase