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.
Mol Pharmacol
2012 Dec 01;826:1042-55. doi: 10.1124/mol.112.080267.
Show Gene links
Show Anatomy links
Locating a plausible binding site for an open-channel blocker, GlyH-101, in the pore of the cystic fibrosis transmembrane conductance regulator.
Norimatsu Y, Ivetac A, Alexander C, O'Donnell N, Frye L, Sansom MS, Dawson DC.
???displayArticle.abstract???
High-throughput screening has led to the identification of small-molecule blockers of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel, but the structural basis of blocker binding remains to be defined. We developed molecular models of the CFTR channel on the basis of homology to the bacterial transporter Sav1866, which could permit blocker binding to be analyzed in silico. The models accurately predicted the existence of a narrow region in the pore that is a likely candidate for the binding site of an open-channel pore blocker such as N-(2-naphthalenyl)-[(3,5-dibromo-2,4-dihydroxyphenyl)methylene]glycine hydrazide (GlyH-101), which is thought to act by entering the channel from the extracellular side. As a more-stringent test of predictions of the CFTR pore model, we applied induced-fit, virtual, ligand-docking techniques to identify potential binding sites for GlyH-101 within the CFTR pore. The highest-scoring docked position was near two pore-lining residues, Phe337 and Thr338, and the rates of reactions of anionic, thiol-directed reagents with cysteines substituted at these positions were slowed in the presence of the blocker, consistent with the predicted repulsive effect of the net negative charge on GlyH-101. When a bulky phenylalanine that forms part of the predicted binding pocket (Phe342) was replaced with alanine, the apparent affinity of the blocker was increased ∼200-fold. A molecular mechanics-generalized Born/surface area analysis of GlyH-101 binding predicted that substitution of Phe342 with alanine would substantially increase blocker affinity, primarily because of decreased intramolecular strain within the blocker-protein complex. This study suggests that GlyH-101 blocks the CFTR channel by binding within the pore bottleneck.
Alexander,
Cystic fibrosis transmembrane conductance regulator: using differential reactivity toward channel-permeant and channel-impermeant thiol-reactive probes to test a molecular model for the pore.
2009, Pubmed,
Xenbase
Alexander,
Cystic fibrosis transmembrane conductance regulator: using differential reactivity toward channel-permeant and channel-impermeant thiol-reactive probes to test a molecular model for the pore.
2009,
Pubmed
,
Xenbase Bai,
Dual roles of the sixth transmembrane segment of the CFTR chloride channel in gating and permeation.
2010,
Pubmed Beck,
Conformational changes in a pore-lining helix coupled to cystic fibrosis transmembrane conductance regulator channel gating.
2008,
Pubmed
,
Xenbase Bompadre,
CFTR gating I: Characterization of the ATP-dependent gating of a phosphorylation-independent CFTR channel (DeltaR-CFTR).
2005,
Pubmed Boucher,
Airway surface dehydration in cystic fibrosis: pathogenesis and therapy.
2007,
Pubmed Cai,
Voltage-dependent gating of the cystic fibrosis transmembrane conductance regulator Cl- channel.
2003,
Pubmed Cai,
Inhibition of heterologously expressed cystic fibrosis transmembrane conductance regulator Cl- channels by non-sulphonylurea hypoglycaemic agents.
1999,
Pubmed Carra,
Theoretical prediction of the binding free energy for mutants of replication protein A.
2012,
Pubmed Csanády,
Strict coupling between CFTR's catalytic cycle and gating of its Cl- ion pore revealed by distributions of open channel burst durations.
2010,
Pubmed
,
Xenbase Cui,
Mutations at arginine 352 alter the pore architecture of CFTR.
2008,
Pubmed
,
Xenbase Cui,
Differential contribution of TM6 and TM12 to the pore of CFTR identified by three sulfonylurea-based blockers.
2012,
Pubmed
,
Xenbase Dailey,
Structure-based drug design: from nucleic acid to membrane protein targets.
2009,
Pubmed Dalton,
New model of cystic fibrosis transmembrane conductance regulator proposes active channel-like conformation.
2012,
Pubmed de Hostos,
Developing novel antisecretory drugs to treat infectious diarrhea.
2011,
Pubmed del Camino,
Blocker protection in the pore of a voltage-gated K+ channel and its structural implications.
2000,
Pubmed Fatehi,
State-dependent access of anions to the cystic fibrosis transmembrane conductance regulator chloride channel pore.
2008,
Pubmed Fischer,
CFTR displays voltage dependence and two gating modes during stimulation.
1994,
Pubmed Friesner,
Extra precision glide: docking and scoring incorporating a model of hydrophobic enclosure for protein-ligand complexes.
2006,
Pubmed Guimarães,
MM-GB/SA rescoring of docking poses in structure-based lead optimization.
2008,
Pubmed Holmgren,
On the use of thiol-modifying agents to determine channel topology.
1996,
Pubmed
,
Xenbase Hwang,
Molecular pharmacology of the CFTR Cl- channel.
1999,
Pubmed Karlin,
Substituted-cysteine accessibility method.
1998,
Pubmed Lê-Quôc,
Evidence for the existence of two classes of sulfhydryl groups essential for membrane-bound succinate dehydrogenase activity.
1981,
Pubmed Linsdell,
Location of a common inhibitor binding site in the cytoplasmic vestibule of the cystic fibrosis transmembrane conductance regulator chloride channel pore.
2005,
Pubmed Liu,
CFTR: covalent modification of cysteine-substituted channels expressed in Xenopus oocytes shows that activation is due to the opening of channels resident in the plasma membrane.
2001,
Pubmed
,
Xenbase Liu,
CFTR: a cysteine at position 338 in TM6 senses a positive electrostatic potential in the pore.
2004,
Pubmed
,
Xenbase Liu,
Variable reactivity of an engineered cysteine at position 338 in cystic fibrosis transmembrane conductance regulator reflects different chemical states of the thiol.
2006,
Pubmed
,
Xenbase Liu,
A possible role for intracellular GSH in spontaneous reaction of a cysteine (T338C) engineered into the Cystic Fibrosis Transmembrane Conductance Regulator.
2008,
Pubmed
,
Xenbase Mansoura,
Cystic fibrosis transmembrane conductance regulator (CFTR) anion binding as a probe of the pore.
1998,
Pubmed
,
Xenbase Mense,
In vivo phosphorylation of CFTR promotes formation of a nucleotide-binding domain heterodimer.
2006,
Pubmed
,
Xenbase Meyer,
Interactions with aromatic rings in chemical and biological recognition.
2003,
Pubmed Mornon,
Molecular models of the open and closed states of the whole human CFTR protein.
2009,
Pubmed Muanprasat,
Discovery of glycine hydrazide pore-occluding CFTR inhibitors: mechanism, structure-activity analysis, and in vivo efficacy.
2004,
Pubmed Norimatsu,
Cystic fibrosis transmembrane conductance regulator: a molecular model defines the architecture of the anion conduction path and locates a "bottleneck" in the pore.
2012,
Pubmed
,
Xenbase Parker,
Analysis of the binding of hydroxamic acid and carboxylic acid inhibitors to the stromelysin-1 (matrix metalloproteinase-3) catalytic domain by isothermal titration calorimetry.
1999,
Pubmed Paton,
Hydrogen bonding and pi-stacking: how reliable are force fields? A critical evaluation of force field descriptions of nonbonded interactions.
2009,
Pubmed Rao,
Improving database enrichment through ensemble docking.
2008,
Pubmed Riordan,
CFTR function and prospects for therapy.
2008,
Pubmed Sanchez,
Cholera toxin - a foe & a friend.
2011,
Pubmed Serrano,
CFTR: Ligand exchange between a permeant anion ([Au(CN)2]-) and an engineered cysteine (T338C) blocks the pore.
2006,
Pubmed
,
Xenbase Sheppard,
CFTR channel pharmacology: novel pore blockers identified by high-throughput screening.
2004,
Pubmed Sheppard,
Mechanism of glibenclamide inhibition of cystic fibrosis transmembrane conductance regulator Cl- channels expressed in a murine cell line.
1997,
Pubmed Sherman,
Use of an induced fit receptor structure in virtual screening.
2006,
Pubmed Shoichet,
Structure-based drug screening for G-protein-coupled receptors.
2012,
Pubmed Smith,
CFTR: covalent and noncovalent modification suggests a role for fixed charges in anion conduction.
2001,
Pubmed
,
Xenbase Sonawane,
Luminally active, nonabsorbable CFTR inhibitors as potential therapy to reduce intestinal fluid loss in cholera.
2006,
Pubmed Sonawane,
Lectin conjugates as potent, nonabsorbable CFTR inhibitors for reducing intestinal fluid secretion in cholera.
2007,
Pubmed Tikhonov,
Voltage dependence of open channel blockade: onset and offset rates.
1998,
Pubmed Villoutreix,
Structure-based virtual ligand screening: recent success stories.
2009,
Pubmed Wang,
Automatic atom type and bond type perception in molecular mechanical calculations.
2006,
Pubmed Wang,
Alternating access to the transmembrane domain of the ATP-binding cassette protein cystic fibrosis transmembrane conductance regulator (ABCC7).
2012,
Pubmed Ward,
Flexibility in the ABC transporter MsbA: Alternating access with a twist.
2007,
Pubmed Woodhull,
Ionic blockage of sodium channels in nerve.
1973,
Pubmed Yang,
Small-molecule CFTR inhibitors slow cyst growth in polycystic kidney disease.
2008,
Pubmed Zhang,
State-dependent chemical reactivity of an engineered cysteine reveals conformational changes in the outer vestibule of the cystic fibrosis transmembrane conductance regulator.
2005,
Pubmed Zhang,
Steady-state interactions of glibenclamide with CFTR: evidence for multiple sites in the pore.
2004,
Pubmed
,
Xenbase Zhang,
Time-dependent interactions of glibenclamide with CFTR: kinetically complex block of macroscopic currents.
2004,
Pubmed
,
Xenbase Zhou,
Probing an open CFTR pore with organic anion blockers.
2002,
Pubmed Zhou,
Direct and indirect effects of mutations at the outer mouth of the cystic fibrosis transmembrane conductance regulator chloride channel pore.
2007,
Pubmed Zhou,
Regulation of conductance by the number of fixed positive charges in the intracellular vestibule of the CFTR chloride channel pore.
2010,
Pubmed
