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.
J Biol Chem
2011 Oct 07;28640:34635-42. doi: 10.1074/jbc.M111.269647.
Show Gene links
Show Anatomy links
Engineering a prokaryotic Cys-loop receptor with a third functional domain.
Goyal R
,
Salahudeen AA
,
Jansen M
.
???displayArticle.abstract???
Prokaryotic members of the Cys-loop receptor ligand-gated ion channel superfamily were recently identified. Previously, Cys-loop receptors were only known from multicellular organisms (metazoans). Contrary to the metazoan Cys-loop receptors, the prokaryotic ones consist of an extracellular (ECD) and a transmembrane domain (TMD), lacking the large intracellular domain (ICD) present in metazoa (between transmembrane segments M3 and M4). Using a chimera approach, we added the 115-amino acid ICD from mammalian serotonin type 3A receptors (5-HT(3A)) to the prokaryotic proton-activated Gloeobacter violaceus ligand-gated ion channel (GLIC). We created 12 GLIC-5-HT(3A)-ICD chimeras by replacing a variable number of amino acids in the short GLIC M3M4 linker with the entire 5-HT(3A)-ICD. Two-electrode voltage clamp recordings after expression in Xenopus laevis oocytes showed that only two chimeras were functional and produced currents upon acidification. The pH(50) was comparable with wild-type GLIC. 5-HT(3A) receptor expression can be inhibited by the chaperone protein RIC-3. We have shown previously that the 5-HT(3A)-ICD is required for the attenuation of 5-HT-induced currents when RIC-3 is co-expressed with 5-HT(3A) receptors in X. laevis oocytes. Expression of both functional 5-HT(3A) chimeras was inhibited by RIC-3 co-expression, indicating appropriate folding of the 5-HT(3A)-ICD in the chimeras. Our results indicate that the ICD can be considered a separate domain that can be removed from or added to the ECD and TMD while maintaining the overall structure and function of the ECD and TMD.
Akk,
Structural elements near the C-terminus are responsible for changes in nicotinic receptor gating kinetics following patch excision.
2000, Pubmed
Akk,
Structural elements near the C-terminus are responsible for changes in nicotinic receptor gating kinetics following patch excision.
2000,
Pubmed
Bali,
The location of a closed channel gate in the GABAA receptor channel.
2007,
Pubmed
,
Xenbase
Bali,
GABA-induced intersubunit conformational movement in the GABAA receptor alpha 1M1-beta 2M3 transmembrane subunit interface: experimental basis for homology modeling of an intravenous anesthetic binding site.
2009,
Pubmed
,
Xenbase
Belelli,
Cloning and functional expression of a human 5-hydroxytryptamine type 3AS receptor subunit.
1995,
Pubmed
,
Xenbase
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
Bouzat,
Coupling of agonist binding to channel gating in an ACh-binding protein linked to an ion channel.
2004,
Pubmed
Bouzat,
Structural basis of the different gating kinetics of fetal and adult acetylcholine receptors.
1994,
Pubmed
Brejc,
Crystal structure of an ACh-binding protein reveals the ligand-binding domain of nicotinic receptors.
2001,
Pubmed
Carland,
Characterization of the effects of charged residues in the intracellular loop on ion permeation in alpha1 glycine receptor channels.
2009,
Pubmed
Castelán,
Cytoplasmic regions adjacent to the M3 and M4 transmembrane segments influence expression and function of alpha7 nicotinic acetylcholine receptors. A study with single amino acid mutants.
2007,
Pubmed
,
Xenbase
Castillo,
Dual role of the RIC-3 protein in trafficking of serotonin and nicotinic acetylcholine receptors.
2005,
Pubmed
,
Xenbase
Celie,
Nicotine and carbamylcholine binding to nicotinic acetylcholine receptors as studied in AChBP crystal structures.
2004,
Pubmed
Charych,
The brefeldin A-inhibited GDP/GTP exchange factor 2, a protein involved in vesicular trafficking, interacts with the beta subunits of the GABA receptors.
2004,
Pubmed
Cherezov,
High-resolution crystal structure of an engineered human beta2-adrenergic G protein-coupled receptor.
2007,
Pubmed
Chien,
Structure of the human dopamine D3 receptor in complex with a D2/D3 selective antagonist.
2010,
Pubmed
Dellisanti,
Crystal structure of the extracellular domain of nAChR alpha1 bound to alpha-bungarotoxin at 1.94 A resolution.
2007,
Pubmed
Duret,
Functional prokaryotic-eukaryotic chimera from the pentameric ligand-gated ion channel family.
2011,
Pubmed
,
Xenbase
Eiselé,
Chimaeric nicotinic-serotonergic receptor combines distinct ligand binding and channel specificities.
1993,
Pubmed
,
Xenbase
Gensler,
Assembly and clustering of acetylcholine receptors containing GFP-tagged epsilon or gamma subunits: selective targeting to the neuromuscular junction in vivo.
2001,
Pubmed
,
Xenbase
Gonzalez-Gutierrez,
Bridging the gap between structural models of nicotinic receptor superfamily ion channels and their corresponding functional states.
2010,
Pubmed
Grenningloh,
The strychnine-binding subunit of the glycine receptor shows homology with nicotinic acetylcholine receptors.
,
Pubmed
Grenningloh,
Glycine vs GABA receptors.
,
Pubmed
Grutter,
A chimera encoding the fusion of an acetylcholine-binding protein to an ion channel is stabilized in a state close to the desensitized form of ligand-gated ion channels.
2005,
Pubmed
Hales,
Common determinants of single channel conductance within the large cytoplasmic loop of 5-hydroxytryptamine type 3 and alpha4beta2 nicotinic acetylcholine receptors.
2006,
Pubmed
Halevi,
Conservation within the RIC-3 gene family. Effectors of mammalian nicotinic acetylcholine receptor expression.
2003,
Pubmed
,
Xenbase
Halevi,
The C. elegans ric-3 gene is required for maturation of nicotinic acetylcholine receptors.
2002,
Pubmed
,
Xenbase
Hibbs,
Principles of activation and permeation in an anion-selective Cys-loop receptor.
2011,
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
Hilf,
Structural basis of open channel block in a prokaryotic pentameric ligand-gated ion channel.
2010,
Pubmed
,
Xenbase
Ilegems,
Noninvasive imaging of 5-HT3 receptor trafficking in live cells: from biosynthesis to endocytosis.
2004,
Pubmed
Ilegems,
Ligand binding transmits conformational changes across the membrane-spanning region to the intracellular side of the 5-HT3 serotonin receptor.
2005,
Pubmed
Jaakola,
The 2.6 angstrom crystal structure of a human A2A adenosine receptor bound to an antagonist.
2008,
Pubmed
Janovjak,
A light-gated, potassium-selective glutamate receptor for the optical inhibition of neuronal firing.
2010,
Pubmed
Jansen,
State-dependent cross-linking of the M2 and M3 segments: functional basis for the alignment of GABAA and acetylcholine receptor M3 segments.
2006,
Pubmed
,
Xenbase
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
Jespersen,
Dual-function vector for protein expression in both mammalian cells and Xenopus laevis oocytes.
2002,
Pubmed
,
Xenbase
Keller,
Adjacent basic amino acid residues recognized by the COP I complex and ubiquitination govern endoplasmic reticulum to cell surface trafficking of the nicotinic acetylcholine receptor alpha-Subunit.
2001,
Pubmed
Kelley,
A cytoplasmic region determines single-channel conductance in 5-HT3 receptors.
2003,
Pubmed
Kobe,
Crystal structure of human T cell leukemia virus type 1 gp21 ectodomain crystallized as a maltose-binding protein chimera reveals structural evolution of retroviral transmembrane proteins.
1999,
Pubmed
Kottwitz,
Intracellular domains of the delta-subunits of Torpedo and rat acetylcholine receptors--expression, purification, and characterization.
2004,
Pubmed
Kukhtina,
Intracellular domain of nicotinic acetylcholine receptor: the importance of being unfolded.
2006,
Pubmed
Kunkel,
Rapid and efficient site-specific mutagenesis without phenotypic selection.
1985,
Pubmed
Li,
Identification of a GABAA receptor anesthetic binding site at subunit interfaces by photolabeling with an etomidate analog.
2006,
Pubmed
Li,
Numerous classes of general anesthetics inhibit etomidate binding to gamma-aminobutyric acid type A (GABAA) receptors.
2010,
Pubmed
Li,
Early fluorescence signals detect transitions at mammalian serotonin transporters.
2002,
Pubmed
,
Xenbase
Long,
Atomic structure of a voltage-dependent K+ channel in a lipid membrane-like environment.
2007,
Pubmed
Meyer,
Identification of a gephyrin binding motif on the glycine receptor beta subunit.
1995,
Pubmed
Mihic,
Sites of alcohol and volatile anaesthetic action on GABA(A) and glycine receptors.
1997,
Pubmed
,
Xenbase
Milone,
Mode switching kinetics produced by a naturally occurring mutation in the cytoplasmic loop of the human acetylcholine receptor epsilon subunit.
1998,
Pubmed
Miyake,
Molecular cloning of human 5-hydroxytryptamine3 receptor: heterogeneity in distribution and function among species.
1995,
Pubmed
Nishida,
Crystal structure of a Kir3.1-prokaryotic Kir channel chimera.
2007,
Pubmed
Nury,
X-ray structures of general anaesthetics bound to a pentameric ligand-gated ion channel.
2011,
Pubmed
Parikh,
Structure of the M2 transmembrane segment of GLIC, a prokaryotic Cys loop receptor homologue from Gloeobacter violaceus, probed by substituted cysteine accessibility.
2011,
Pubmed
Peters,
The 5-hydroxytryptamine type 3 (5-HT3) receptor reveals a novel determinant of single-channel conductance.
2004,
Pubmed
Ren,
Structural determinants of alpha4beta2 nicotinic acetylcholine receptor trafficking.
2005,
Pubmed
Rosenbaum,
GPCR engineering yields high-resolution structural insights into beta2-adrenergic receptor function.
2007,
Pubmed
Sadtler,
A basic cluster determines topology of the cytoplasmic M3-M4 loop of the glycine receptor alpha1 subunit.
2003,
Pubmed
,
Xenbase
Schofield,
Sequence and functional expression of the GABA A receptor shows a ligand-gated receptor super-family.
,
Pubmed
,
Xenbase
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
Unwin,
Refined structure of the nicotinic acetylcholine receptor at 4A resolution.
2005,
Pubmed
Valor,
Role of the large cytoplasmic loop of the alpha 7 neuronal nicotinic acetylcholine receptor subunit in receptor expression and function.
2002,
Pubmed
,
Xenbase
Wang,
GABA(A)-receptor-associated protein links GABA(A) receptors and the cytoskeleton.
1999,
Pubmed
Williams,
Ric-3 promotes functional expression of the nicotinic acetylcholine receptor alpha7 subunit in mammalian cells.
2005,
Pubmed
,
Xenbase
Wu,
Structures of the CXCR4 chemokine GPCR with small-molecule and cyclic peptide antagonists.
2010,
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
