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Channels (Austin)
2012 Jan 01;62:111-23. doi: 10.4161/chan.19540.
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Tryptophan scanning mutagenesis reveals distortions in the helical structure of the δM4 transmembrane domain of the Torpedo californica nicotinic acetylcholine receptor.
Caballero-Rivera D
,
Cruz-Nieves OA
,
Oyola-Cintrón J
,
Torres-Nunez DA
,
Otero-Cruz JD
,
Lasalde-Dominicci JA
.
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The lipid-protein interface is an important domain of the nicotinic acetylcholine receptor (nAChR) that has recently garnered increased relevance. Several studies have made significant advances toward determining the structure and dynamics of the lipid-exposed domains of the nAChR. However, there is still a need to gain insight into the mechanism by which lipid-protein interactions regulate the function and conformational transitions of the nAChR. In this study, we extended the tryptophan scanning mutagenesis (TrpScanM) approach to dissect secondary structure and monitor the conformational changes experienced by the δM4 transmembrane domain (TMD) of the Torpedo californica nAChR, and to identify which positions on this domain are potentially linked to the regulation of ion channel kinetics. The difference in oscillation patterns between the closed- and open-channel states suggests a substantial conformational change along this domain as a consequence of channel activation. Furthermore, TrpScanM revealed distortions along the helical structure of this TMD that are not present on current models of the nAChR. Our results show that a Thr-Pro motif at positions 462-463 markedly bends the helical structure of the TMD, consistent with the recent crystallographic structure of the GluCl Cys-loop receptor which reveals a highly bent TMD4 in each subunit. This Thr-Pro motif acts as a molecular hinge that delineates two gating blocks in the δM4 TMD. These results suggest a model in which a hinge-bending motion that tilts the helical structure is combined with a spring-like motion during transition between the closed- and open-channel states of the δM4 TMD.
Baenziger,
Fourier transform infrared and hydrogen/deuterium exchange reveal an exchange-resistant core of alpha-helical peptide hydrogens in the nicotinic acetylcholine receptor.
1995, Pubmed
Baenziger,
Fourier transform infrared and hydrogen/deuterium exchange reveal an exchange-resistant core of alpha-helical peptide hydrogens in the nicotinic acetylcholine receptor.
1995,
Pubmed
Báez-Pagán,
Potential role of caveolin-1-positive domains in the regulation of the acetylcholine receptor's activatable pool: implications in the pathogenesis of a novel congenital myasthenic syndrome.
2008,
Pubmed
,
Xenbase
Bansal,
HELANAL: a program to characterize helix geometry in proteins.
2000,
Pubmed
Barlow,
Helix geometry in proteins.
1988,
Pubmed
Barrantes,
Structural basis for lipid modulation of nicotinic acetylcholine receptor function.
2004,
Pubmed
Blanton,
Identifying the lipid-protein interface of the Torpedo nicotinic acetylcholine receptor: secondary structure implications.
1994,
Pubmed
Blanton,
Probing the structure of the nicotinic acetylcholine receptor ion channel with the uncharged photoactivable compound -3H-diazofluorene.
1998,
Pubmed
Blanton,
Mapping the lipid-exposed regions in the Torpedo californica nicotinic acetylcholine receptor.
1992,
Pubmed
Bocquet,
X-ray structure of a pentameric ligand-gated ion channel in an apparently open conformation.
2009,
Pubmed
Bouzat,
Nicotinic receptor fourth transmembrane domain: hydrogen bonding by conserved threonine contributes to channel gating kinetics.
2000,
Pubmed
Bouzat,
Subunit-selective contribution to channel gating of the M4 domain of the nicotinic receptor.
2002,
Pubmed
Brandl,
Hypothesis about the function of membrane-buried proline residues in transport proteins.
1986,
Pubmed
Brannigan,
Embedded cholesterol in the nicotinic acetylcholine receptor.
2008,
Pubmed
Brejc,
Crystal structure of an ACh-binding protein reveals the ligand-binding domain of nicotinic receptors.
2001,
Pubmed
Caballero-Rivera,
Fourier transform coupled tryptophan scanning mutagenesis identifies a bending point on the lipid-exposed δM3 transmembrane domain of the Torpedo californica nicotinic acetylcholine receptor.
2011,
Pubmed
,
Xenbase
Carman,
Regulation of G protein-coupled receptor kinases by caveolin.
1999,
Pubmed
Cha,
Atomic scale movement of the voltage-sensing region in a potassium channel measured via spectroscopy.
1999,
Pubmed
Choe,
Three distinct structural environments of a transmembrane domain in the inwardly rectifying potassium channel ROMK1 defined by perturbation.
1995,
Pubmed
,
Xenbase
Chothia,
Structural invariants in protein folding.
1975,
Pubmed
Collins,
Scanning mutagenesis of the putative transmembrane segments of Kir2.1, an inward rectifier potassium channel.
1997,
Pubmed
,
Xenbase
Cordes,
Proline-induced distortions of transmembrane helices.
2002,
Pubmed
Corringer,
Nicotinic receptors at the amino acid level.
2000,
Pubmed
Couet,
Identification of peptide and protein ligands for the caveolin-scaffolding domain. Implications for the interaction of caveolin with caveolae-associated proteins.
1997,
Pubmed
Cruz-Martín,
Tryptophan substitutions at lipid-exposed positions of the gamma M3 transmembrane domain increase the macroscopic ionic current response of the Torpedo californica nicotinic acetylcholine receptor.
2001,
Pubmed
,
Xenbase
Cukras,
Structural and functional determinants of conserved lipid interaction domains of inward rectifying Kir6.2 channels.
2002,
Pubmed
Dellisanti,
Crystal structure of the extracellular domain of nAChR alpha1 bound to alpha-bungarotoxin at 1.94 A resolution.
2007,
Pubmed
Deupi,
Influence of the g- conformation of Ser and Thr on the structure of transmembrane helices.
2010,
Pubmed
Deupi,
Ser and Thr residues modulate the conformation of pro-kinked transmembrane alpha-helices.
2004,
Pubmed
Díaz-De León,
Tryptophan scanning of the acetylcholine receptor's betaM4 transmembrane domain: decoding allosteric linkage at the lipid-protein interface with ion-channel gating.
2008,
Pubmed
Gether,
Uncovering molecular mechanisms involved in activation of G protein-coupled receptors.
2000,
Pubmed
Glauner,
Spectroscopic mapping of voltage sensor movement in the Shaker potassium channel.
1999,
Pubmed
,
Xenbase
Guzmán,
Tryptophan scanning mutagenesis in the alphaM3 transmembrane domain of the Torpedo californica acetylcholine receptor: functional and structural implications.
2003,
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
Honse,
Sites in the fourth membrane-associated domain regulate alcohol sensitivity of the NMDA receptor.
2004,
Pubmed
Irizarry,
Opening the KcsA K+ channel: tryptophan scanning and complementation analysis lead to mutants with altered gating.
2002,
Pubmed
Jenkins,
Tryptophan scanning mutagenesis in TM4 of the GABA(A) receptor alpha1 subunit: implications for modulation by inhaled anesthetics and ion channel structure.
2002,
Pubmed
Kalamida,
Muscle and neuronal nicotinic acetylcholine receptors. Structure, function and pathogenicity.
2007,
Pubmed
Karlin,
Emerging structure of the nicotinic acetylcholine receptors.
2002,
Pubmed
Kerr,
Molecular dynamics simulations of isolated transmembrane helices of potassium channels.
1996,
Pubmed
Lasalde,
Tryptophan substitutions at the lipid-exposed transmembrane segment M4 of Torpedo californica acetylcholine receptor govern channel gating.
1996,
Pubmed
,
Xenbase
Lee,
Mutations in the M4 domain of Torpedo californica acetylcholine receptor dramatically alter ion channel function.
1994,
Pubmed
,
Xenbase
Li,
Functional role of the cysteine 451 thiol group in the M4 helix of the gamma subunit of Torpedo californica acetylcholine receptor.
1990,
Pubmed
,
Xenbase
Li,
Site-specific mutations of nicotinic acetylcholine receptor at the lipid-protein interface dramatically alter ion channel gating.
1992,
Pubmed
,
Xenbase
Li-Smerin,
Helical structure of the COOH terminus of S3 and its contribution to the gating modifier toxin receptor in voltage-gated ion channels.
2001,
Pubmed
Li-Smerin,
A localized interaction surface for voltage-sensing domains on the pore domain of a K+ channel.
2000,
Pubmed
,
Xenbase
Li-Smerin,
alpha-helical structural elements within the voltage-sensing domains of a K(+) channel.
2000,
Pubmed
,
Xenbase
Luecke,
Structure of bacteriorhodopsin at 1.55 A resolution.
1999,
Pubmed
Lugovskoy,
Spatial structure of the M3 transmembrane segment of the nicotinic acetylcholine receptor alpha subunit.
1998,
Pubmed
Maduke,
A decade of CLC chloride channels: structure, mechanism, and many unsettled questions.
2000,
Pubmed
McGregor,
Analysis of the relationship between side-chain conformation and secondary structure in globular proteins.
1987,
Pubmed
Méthot,
Secondary structure of the exchange-resistant core from the nicotinic acetylcholine receptor probed directly by infrared spectroscopy and hydrogen/deuterium exchange.
1998,
Pubmed
Mitra,
Structural dynamics of the M4 transmembrane segment during acetylcholine receptor gating.
2004,
Pubmed
Miyazawa,
Structure and gating mechanism of the acetylcholine receptor pore.
2003,
Pubmed
Navedo,
Tryptophan substitutions reveal the role of nicotinic acetylcholine receptor alpha-TM3 domain in channel gating: differences between Torpedo and muscle-type AChR.
2004,
Pubmed
,
Xenbase
Nilsson,
Proline-induced disruption of a transmembrane alpha-helix in its natural environment.
1998,
Pubmed
Okamoto,
Caveolins, a family of scaffolding proteins for organizing "preassembled signaling complexes" at the plasma membrane.
1998,
Pubmed
Ortiz-Acevedo,
Tryptophan scanning mutagenesis of the gammaM4 transmembrane domain of the acetylcholine receptor from Torpedo californica.
2004,
Pubmed
,
Xenbase
Ortiz-Miranda,
Mutations in the M4 domain of the Torpedo californica nicotinic acetylcholine receptor alter channel opening and closing.
1997,
Pubmed
,
Xenbase
Otero-Cruz,
Tryptophan-scanning mutagenesis in the alphaM3 transmembrane domain of the muscle-type acetylcholine receptor. A spring model revealed.
2007,
Pubmed
,
Xenbase
Powl,
Identification of the hydrophobic thickness of a membrane protein using fluorescence spectroscopy: studies with the mechanosensitive channel MscL.
2005,
Pubmed
Ri,
The role of a conserved proline residue in mediating conformational changes associated with voltage gating of Cx32 gap junctions.
1999,
Pubmed
,
Xenbase
Sankararamakrishnan,
Characterization of proline-containing alpha-helix (helix F model of bacteriorhodopsin) by molecular dynamics studies.
1993,
Pubmed
Sankararamakrishnan,
Geometry of proline-containing alpha-helices in proteins.
1992,
Pubmed
Sansom,
Hinges, swivels and switches: the role of prolines in signalling via transmembrane alpha-helices.
2000,
Pubmed
Sansom,
Structure and function of channel-forming peptaibols.
1993,
Pubmed
Santiago,
Probing the effects of membrane cholesterol in the Torpedo californica acetylcholine receptor and the novel lipid-exposed mutation alpha C418W in Xenopus oocytes.
2001,
Pubmed
,
Xenbase
Santiago,
Tryptophan scanning mutagenesis in the TM3 domain of the Torpedo californica acetylcholine receptor beta subunit reveals an alpha-helical structure.
2004,
Pubmed
,
Xenbase
Shen,
Slow-channel mutation in acetylcholine receptor alphaM4 domain and its efficient knockdown.
2006,
Pubmed
Silberberg,
Secondary structure and gating rearrangements of transmembrane segments in rat P2X4 receptor channels.
2005,
Pubmed
,
Xenbase
Tamamizu,
Functional effects of periodic tryptophan substitutions in the alpha M4 transmembrane domain of the Torpedo californica nicotinic acetylcholine receptor.
2000,
Pubmed
,
Xenbase
Tamamizu,
Alteration in ion channel function of mouse nicotinic acetylcholine receptor by mutations in the M4 transmembrane domain.
1999,
Pubmed
,
Xenbase
Tieleman,
Proline-induced hinges in transmembrane helices: possible roles in ion channel gating.
2001,
Pubmed
Unwin,
Nicotinic acetylcholine receptor at 9 A resolution.
1993,
Pubmed
Unwin,
Acetylcholine receptor channel imaged in the open state.
1995,
Pubmed
Unwin,
Refined structure of the nicotinic acetylcholine receptor at 4A resolution.
2005,
Pubmed
Wang,
Tryptophan scanning of D1S6 and D4S6 C-termini in voltage-gated sodium channels.
2003,
Pubmed
Williams,
Proline residues in transmembrane helices: structural or dynamic role?
1991,
Pubmed
Woolfson,
The influence of proline residues on alpha-helical structure.
1990,
Pubmed
Xu,
Conformational dynamics of the nicotinic acetylcholine receptor channel: a 35-ns molecular dynamics simulation study.
2005,
Pubmed
Yohannan,
The evolution of transmembrane helix kinks and the structural diversity of G protein-coupled receptors.
2004,
Pubmed
Zhu,
Gating transition of pentameric ligand-gated ion channels.
2009,
Pubmed
