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.
Proc Natl Acad Sci U S A
2013 May 14;11020:8272-7. doi: 10.1073/pnas.1220398110.
Show Gene links
Show Anatomy links
Dynamics of internal pore opening in K(V) channels probed by a fluorescent unnatural amino acid.
Kalstrup T
,
Blunck R
.
???displayArticle.abstract???
Atomic-scale models on the gating mechanism of voltage-gated potassium channels (Kv) are based on linear interpolations between static structures of their initial and final state derived from crystallography and molecular dynamics simulations, and, thus, lack dynamic structural information. The lack of information on dynamics and intermediate states makes it difficult to associate the structural with the dynamic functional data obtained with electrophysiology. Although voltage-clamp fluorometry fills this gap, it is limited to sites extracellularly accessible, when the key region for gating is located at the cytosolic side of the channels. Here, we solved this problem by performing voltage-clamp fluorometry with a fluorescent unnatural amino acid. By using an orthogonal tRNA-synthetase pair, the fluorescent unnatural amino acid was incorporated in the Shaker voltage-gated potassium channel at key regions that were previously inaccessible. Thus, we defined which parts act independently and which parts act cooperatively and found pore opening to occur in two sequential transitions.
Baker,
Three transmembrane conformations and sequence-dependent displacement of the S4 domain in shaker K+ channel gating.
1998, Pubmed
Baker,
Three transmembrane conformations and sequence-dependent displacement of the S4 domain in shaker K+ channel gating.
1998,
Pubmed
Bannister,
Optical detection of rate-determining ion-modulated conformational changes of the ether-à-go-go K+ channel voltage sensor.
2005,
Pubmed
,
Xenbase
Batulan,
An intersubunit interaction between S4-S5 linker and S6 is responsible for the slow off-gating component in Shaker K+ channels.
2010,
Pubmed
Bezanilla,
Inactivation of the sodium channel. I. Sodium current experiments.
1977,
Pubmed
Bezanilla,
Gating of Shaker K+ channels: II. The components of gating currents and a model of channel activation.
1994,
Pubmed
,
Xenbase
Blunck,
Detecting rearrangements of shaker and NaChBac in real-time with fluorescence spectroscopy in patch-clamped mammalian cells.
2004,
Pubmed
,
Xenbase
Blunck,
Mechanism of electromechanical coupling in voltage-gated potassium channels.
2012,
Pubmed
Cha,
Voltage sensors in domains III and IV, but not I and II, are immobilized by Na+ channel fast inactivation.
1999,
Pubmed
,
Xenbase
Cha,
Characterizing voltage-dependent conformational changes in the Shaker K+ channel with fluorescence.
1997,
Pubmed
,
Xenbase
Cha,
Structural implications of fluorescence quenching in the Shaker K+ channel.
1998,
Pubmed
Claydon,
4-aminopyridine prevents the conformational changes associated with p/c-type inactivation in shaker channels.
2007,
Pubmed
,
Xenbase
Claydon,
Voltage clamp fluorimetry studies of mammalian voltage-gated K(+) channel gating.
2007,
Pubmed
COLE,
Potassium ion current in the squid giant axon: dynamic characteristic.
1960,
Pubmed
Dahan,
A fluorophore attached to nicotinic acetylcholine receptor beta M2 detects productive binding of agonist to the alpha delta site.
2004,
Pubmed
,
Xenbase
Ding,
Effect of S6 tail mutations on charge movement in Shaker potassium channels.
2003,
Pubmed
Faure,
A limited 4 Å radial displacement of the S4-S5 linker is sufficient for internal gate closing in Kv channels.
2012,
Pubmed
Freites,
Microscopic origin of gating current fluctuations in a potassium channel voltage sensor.
2012,
Pubmed
Gagnon,
Membrane topology of loop 13-14 of the Na+/glucose cotransporter (SGLT1): a SCAM and fluorescent labelling study.
2005,
Pubmed
,
Xenbase
Gandhi,
Reconstructing voltage sensor-pore interaction from a fluorescence scan of a voltage-gated K+ channel.
2000,
Pubmed
Haddad,
Mode shift of the voltage sensors in Shaker K+ channels is caused by energetic coupling to the pore domain.
2011,
Pubmed
,
Xenbase
Hoshi,
Biophysical and molecular mechanisms of Shaker potassium channel inactivation.
1990,
Pubmed
,
Xenbase
Jensen,
Mechanism of voltage gating in potassium channels.
2012,
Pubmed
Labro,
Kv channel gating requires a compatible S4-S5 linker and bottom part of S6, constrained by non-interacting residues.
2008,
Pubmed
Ledwell,
Mutations in the S4 region isolate the final voltage-dependent cooperative step in potassium channel activation.
1999,
Pubmed
,
Xenbase
Lee,
Genetic incorporation of a small, environmentally sensitive, fluorescent probe into proteins in Saccharomyces cerevisiae.
2009,
Pubmed
Li,
An intermediate state of the gamma-aminobutyric acid transporter GAT1 revealed by simultaneous voltage clamp and fluorescence.
2000,
Pubmed
,
Xenbase
Li,
Early fluorescence signals detect transitions at mammalian serotonin transporters.
2002,
Pubmed
,
Xenbase
Loboda,
Resolving the gating charge movement associated with late transitions in K channel activation.
2001,
Pubmed
Long,
Crystal structure of a mammalian voltage-dependent Shaker family K+ channel.
2005,
Pubmed
Long,
Atomic structure of a voltage-dependent K+ channel in a lipid membrane-like environment.
2007,
Pubmed
Long,
Voltage sensor of Kv1.2: structural basis of electromechanical coupling.
2005,
Pubmed
Lu,
Ion conduction pore is conserved among potassium channels.
2001,
Pubmed
Mannuzzu,
Independence and cooperativity in rearrangements of a potassium channel voltage sensor revealed by single subunit fluorescence.
2000,
Pubmed
,
Xenbase
Mannuzzu,
Direct physical measure of conformational rearrangement underlying potassium channel gating.
1996,
Pubmed
,
Xenbase
McCormack,
A characterization of the activating structural rearrangements in voltage-dependent Shaker K+ channels.
1994,
Pubmed
Meinild,
Fluorescence studies of ligand-induced conformational changes of the Na(+)/glucose cotransporter.
2002,
Pubmed
,
Xenbase
Muroi,
Molecular determinants of coupling between the domain III voltage sensor and pore of a sodium channel.
2010,
Pubmed
Nowak,
Nicotinic receptor binding site probed with unnatural amino acid incorporation in intact cells.
1995,
Pubmed
,
Xenbase
Pantazis,
Relative transmembrane segment rearrangements during BK channel activation resolved by structurally assigned fluorophore-quencher pairing.
2012,
Pubmed
,
Xenbase
Pantoja,
Single-molecule imaging of a fluorescent unnatural amino acid incorporated into nicotinic receptors.
2009,
Pubmed
,
Xenbase
Pathak,
The cooperative voltage sensor motion that gates a potassium channel.
2005,
Pubmed
,
Xenbase
Pless,
Contributions of counter-charge in a potassium channel voltage-sensor domain.
2011,
Pubmed
,
Xenbase
Savalli,
Voltage-dependent conformational changes in human Ca(2+)- and voltage-activated K(+) channel, revealed by voltage-clamp fluorometry.
2006,
Pubmed
,
Xenbase
Schoppa,
Activation of Shaker potassium channels. II. Kinetics of the V2 mutant channel.
1998,
Pubmed
,
Xenbase
Schoppa,
Activation of Shaker potassium channels. III. An activation gating model for wild-type and V2 mutant channels.
1998,
Pubmed
,
Xenbase
Sigg,
Fast gating in the Shaker K+ channel and the energy landscape of activation.
2003,
Pubmed
,
Xenbase
Tao,
A gating charge transfer center in voltage sensors.
2010,
Pubmed
,
Xenbase
Vargas,
In search of a consensus model of the resting state of a voltage-sensing domain.
2011,
Pubmed
Villalba-Galea,
S4-based voltage sensors have three major conformations.
2008,
Pubmed
Yang,
How does the W434F mutation block current in Shaker potassium channels?
1997,
Pubmed
,
Xenbase
Yarov-Yarovoy,
Structural basis for gating charge movement in the voltage sensor of a sodium channel.
2012,
Pubmed
Zagotta,
Shaker potassium channel gating. III: Evaluation of kinetic models for activation.
1994,
Pubmed
,
Xenbase
Zheng,
Gating rearrangements in cyclic nucleotide-gated channels revealed by patch-clamp fluorometry.
2000,
Pubmed
