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.
ACS Omega
2016 Dec 13;16:1205-1219. doi: 10.1021/acsomega.6b00202.
Show Gene links
Show Anatomy links
Structural Heterogeneity of CNGA1 Channels Revealed by Electrophysiology and Single-Molecule Force Spectroscopy.
Maity S
,
Marchesi A
,
Torre V
,
Mazzolini M
.
???displayArticle.abstract???
The determination at atomic resolution of the three-dimensional molecular structure of membrane proteins such as receptors and several ion channels has been a major breakthrough in structural biology. The molecular structure of several members of the superfamily of voltage-gated ionic channels such as K+ and Na+ is now available. However, despite several attempts, the molecular structure at atomic resolution of the full cyclic nucleotide-gated (CNG) ion channel, although a member of the same superfamily of voltage-gated ion channels, has not been obtained yet, neither by X-ray crystallography nor by electron cryomicroscopy (cryo-EM). It is possible that CNG channels have a high structural heterogeneity, making difficult crystallization and single-particle analysis. To address this issue, we have combined single-molecule force spectroscopy (SMFS) and electrophysiological experiments to characterize the structural heterogeneity of CNGA1 channels expressed in Xenopus laevis oocytes. The unfolding of the cytoplasmic domain had force peaks, occurring with a probability from 0.2 to 0.96. Force peaks during the unfolding of the transmembrane domain had a probability close to 1, but the distribution of the increase in contour length between two successive force peaks had multiple maxima differing by tens of nanometers. Concomitant electrophysiological experiments showed that the rundown in mutant channels S399C is highly variable and that the effect of thiol reagents when specific residues were mutated was consistent with a dynamic structural heterogeneity. These results show that CNGA1 channels have a wide spectrum of native conformations that are difficult to detect with X-ray crystallography and cryo-EM.
Akabas,
Acetylcholine receptor channel structure probed in cysteine-substitution mutants.
1992,
Pubmed
,
Xenbase
Anderson,
Phylogeny of ion channels: clues to structure and function.
2001,
Pubmed
Arcangeletti,
Multiple mechanisms underlying rectification in retinal cyclic nucleotide-gated (CNGA1) channels.
2013,
Pubmed
,
Xenbase
Arinaminpathy,
Molecular dynamics simulations of the ligand-binding domain of the ionotropic glutamate receptor GluR2.
2002,
Pubmed
Becchetti,
Cyclic nucleotide-gated channels: intra- and extracellular accessibility to Cd2+ of substituted cysteine residues within the P-loop.
2000,
Pubmed
,
Xenbase
Bell,
Models for the specific adhesion of cells to cells.
1978,
Pubmed
Bosshart,
The transmembrane protein KpOmpA anchoring the outer membrane of Klebsiella pneumoniae unfolds and refolds in response to tensile load.
2012,
Pubmed
Brams,
Family of prokaryote cyclic nucleotide-modulated ion channels.
2014,
Pubmed
,
Xenbase
Bustamante,
Entropic elasticity of lambda-phage DNA.
1994,
Pubmed
Bénitah,
Adjacent pore-lining residues within sodium channels identified by paired cysteine mutagenesis.
1996,
Pubmed
,
Xenbase
Craven,
CNG and HCN channels: two peas, one pod.
2006,
Pubmed
Derebe,
Structural studies of ion permeation and Ca2+ blockage of a bacterial channel mimicking the cyclic nucleotide-gated channel pore.
2011,
Pubmed
Domene,
Potassium channel, ions, and water: simulation studies based on the high resolution X-ray structure of KcsA.
2003,
Pubmed
Evans,
Dynamic strength of molecular adhesion bonds.
1997,
Pubmed
Ge,
Locating an extracellular K+-dependent interaction site that modulates betaine-binding of the Na+-coupled betaine symporter BetP.
2011,
Pubmed
Glusker,
Structural aspects of metal liganding to functional groups in proteins.
1991,
Pubmed
Gorostiza,
Molecular handles for the mechanical manipulation of single-membrane proteins in living cells.
2005,
Pubmed
,
Xenbase
Hastrup,
Symmetrical dimer of the human dopamine transporter revealed by cross-linking Cys-306 at the extracellular end of the sixth transmembrane segment.
2001,
Pubmed
Higgins,
Molecular architecture of a retinal cGMP-gated channel: the arrangement of the cytoplasmic domains.
2002,
Pubmed
Holmgren,
The activation gate of a voltage-gated K+ channel can be trapped in the open state by an intersubunit metal bridge.
1998,
Pubmed
Huynh,
Structure of the full-length TRPV2 channel by cryo-EM.
2016,
Pubmed
Jan,
A superfamily of ion channels.
1990,
Pubmed
Jiang,
The open pore conformation of potassium channels.
2002,
Pubmed
Jiang,
X-ray structure of a voltage-dependent K+ channel.
2003,
Pubmed
Karlin,
Substituted-cysteine accessibility method.
1998,
Pubmed
Kaupp,
Primary structure and functional expression from complementary DNA of the rod photoreceptor cyclic GMP-gated channel.
1989,
Pubmed
,
Xenbase
Kaupp,
Cyclic nucleotide-gated ion channels.
2002,
Pubmed
Kawamura,
Kinetic, energetic, and mechanical differences between dark-state rhodopsin and opsin.
2013,
Pubmed
Kedrov,
Differentiating ligand and inhibitor interactions of a single antiporter.
2006,
Pubmed
Kotamarthi,
Multiple unfolding pathways of leucine binding protein (LBP) probed by single-molecule force spectroscopy (SMFS).
2013,
Pubmed
Kowal,
Ligand-induced structural changes in the cyclic nucleotide-modulated potassium channel MloK1.
2014,
Pubmed
Laio,
Physical origin of selectivity in ionic channels of biological membranes.
1999,
Pubmed
Liao,
Structure of the TRPV1 ion channel determined by electron cryo-microscopy.
2013,
Pubmed
Lolicato,
Tetramerization dynamics of C-terminal domain underlies isoform-specific cAMP gating in hyperpolarization-activated cyclic nucleotide-gated channels.
2011,
Pubmed
,
Xenbase
Long,
Crystal structure of a mammalian voltage-dependent Shaker family K+ channel.
2005,
Pubmed
Macdonald,
Protein-protein interactions affect alpha helix stability in crowded environments.
2015,
Pubmed
Maity,
Conformational rearrangements in the transmembrane domain of CNGA1 channels revealed by single-molecule force spectroscopy.
2015,
Pubmed
,
Xenbase
Manglik,
Structural Insights into the Dynamic Process of β2-Adrenergic Receptor Signaling.
2015,
Pubmed
Marchesi,
Gating of cyclic nucleotide-gated channels is voltage dependent.
2012,
Pubmed
Matulef,
Multimerization of the ligand binding domains of cyclic nucleotide-gated channels.
2002,
Pubmed
,
Xenbase
Mazzolini,
Movement of the C-helix during the gating of cyclic nucleotide-gated channels.
2002,
Pubmed
,
Xenbase
Mazzolini,
A comparison of electrophysiological properties of the CNGA1, CNGA1tandem and CNGA1cys-free channels.
2008,
Pubmed
,
Xenbase
Mazzolini,
The analysis of desensitizing CNGA1 channels reveals molecular interactions essential for normal gating.
2009,
Pubmed
,
Xenbase
Mazzolini,
Gating in CNGA1 channels.
2010,
Pubmed
Nair,
Movements of native C505 during channel gating in CNGA1 channels.
2009,
Pubmed
Nair,
Conformational rearrangements in the S6 domain and C-linker during gating in CNGA1 channels.
2009,
Pubmed
Napolitano,
A structural, functional, and computational analysis suggests pore flexibility as the base for the poor selectivity of CNG channels.
2015,
Pubmed
,
Xenbase
Nygaard,
The dynamic process of β(2)-adrenergic receptor activation.
2013,
Pubmed
Oesterhelt,
Unfolding pathways of individual bacteriorhodopsins.
2000,
Pubmed
Park,
Dynamic single-molecule force spectroscopy of rhodopsin in native membranes.
2015,
Pubmed
Payandeh,
The crystal structure of a voltage-gated sodium channel.
2011,
Pubmed
Peng,
Atomic force microscopy reveals parallel mechanical unfolding pathways of T4 lysozyme: evidence for a kinetic partitioning mechanism.
2008,
Pubmed
Rief,
Reversible unfolding of individual titin immunoglobulin domains by AFM.
1997,
Pubmed
Rosenbaum,
Dissecting intersubunit contacts in cyclic nucleotide-gated ion channels.
2002,
Pubmed
,
Xenbase
Santacroce,
Imaging of Xenopus laevis oocyte plasma membrane in physiological-like conditions by atomic force microscopy.
2013,
Pubmed
,
Xenbase
Schünke,
Structural insights into conformational changes of a cyclic nucleotide-binding domain in solution from Mesorhizobium loti K1 channel.
2011,
Pubmed
Tanuj Sapra,
Detecting molecular interactions that stabilize native bovine rhodopsin.
2006,
Pubmed
Vinothkumar,
Membrane protein structures without crystals, by single particle electron cryomicroscopy.
2015,
Pubmed
Yarov-Yarovoy,
Structural basis for gating charge movement in the voltage sensor of a sodium channel.
2012,
Pubmed
Yu,
Overview of molecular relationships in the voltage-gated ion channel superfamily.
2005,
Pubmed
Zagotta,
Structural basis for modulation and agonist specificity of HCN pacemaker channels.
2003,
Pubmed
Zocher,
Cholesterol increases kinetic, energetic, and mechanical stability of the human β2-adrenergic receptor.
2012,
Pubmed
Zubcevic,
Cryo-electron microscopy structure of the TRPV2 ion channel.
2016,
Pubmed