Proline scan of the HERG channel S6 helix reveals the location of the intracellular pore gate.

Abstract:

:In Shaker-like channels, the activation gate is formed at the bundle crossing by the convergence of the inner S6 helices near a conserved proline-valine-proline motif, which introduces a kink that allows for electromechanical coupling with voltage sensor motions via the S4-S5 linker. Human ether-a-go-go-related gene (hERG) channels lack the proline-valine-proline motif and the location of the intracellular pore gate and how it is coupled to S4 movement is less clear. Here, we show that proline substitutions within the S6 of hERG perturbed pore gate closure, trapping channels in the open state. Performing a proline scan of the inner S6 helix, from Ile(655) to Tyr(667) revealed that gate perturbation occurred with proximal (I655P-Q664P), but not distal (R665P-Y667P) substitutions, suggesting that Gln(664) marks the position of the intracellular gate in hERG channels. Using voltage-clamp fluorimetry and gating current analysis, we demonstrate that proline substitutions trap the activation gate open by disrupting the coupling between the voltage-sensing unit and the pore of the channel. We characterize voltage sensor movement in one such trapped-open mutant channel and demonstrate the kinetics of what we interpret to be intrinsic hERG voltage sensor movement.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Thouta S,Sokolov S,Abe Y,Clark SJ,Cheng YM,Claydon TW

doi

10.1016/j.bpj.2014.01.035

subject

Has Abstract

pub_date

2014-03-04 00:00:00

pages

1057-69

issue

5

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(14)00137-4

journal_volume

106

pub_type

杂志文章
  • A mesoscale model of DNA and its renaturation.

    abstract::A mesoscale model of DNA is presented (3SPN.1), extending the scheme previously developed by our group. Each nucleotide is mapped onto three interaction sites. Solvent is accounted for implicitly through a medium-effective dielectric constant and electrostatic interactions are treated at the level of Debye-Hückel theo...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2008.09.061

    authors: Sambriski EJ,Schwartz DC,de Pablo JJ

    更新日期:2009-03-04 00:00:00

  • A mathematical model of the slow force response to stretch in rat ventricular myocytes.

    abstract::We developed a model of the rat ventricular myocyte at room temperature to predict the relative effects of different mechanisms on the cause of the slow increase in force in response to a step change in muscle length. We performed simulations in the presence of stretch-dependent increases in flux through the Na(+)-H(+...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.106.095463

    authors: Niederer SA,Smith NP

    更新日期:2007-06-01 00:00:00

  • A theory of thermal fluctuations in DNA miniplasmids.

    abstract::A recent analysis of the normal modes of vibration of a ring formed by bringing together and sealing, with or without the addition of twist, the ends of rods that are straight when stress free is taken as the basis for a theory of the statistical thermodynamics of a canonical ensemble of DNA minicircles with specified...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(98)77962-7

    authors: Tobias I

    更新日期:1998-05-01 00:00:00

  • Calcium-Dependent Structural Dynamics of a Spin-Labeled RyR Peptide Bound to Calmodulin.

    abstract::We have used chemical synthesis, electron paramagnetic resonance (EPR), and circular dichroism to detect and analyze the structural dynamics of a ryanodine receptor (RyR) peptide bound to calmodulin (CaM). The skeletal muscle calcium release channel RyR1 is activated by Ca2+-free CaM and inhibited by Ca2+-bound CaM. T...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2016.10.025

    authors: Her C,McCaffrey JE,Thomas DD,Karim CB

    更新日期:2016-12-06 00:00:00

  • Osmotic and pH transmembrane gradients control the lytic power of melittin.

    abstract::Transmembrane osmotic gradients applied on large unilamellar 1-palmitoyl-2-oleoyl-phosphatidylcholine vesicles were used to modulate the potency of melittin to induce leakage. Melittin, an amphipathic peptide, changes the permeability of vesicles, as studied using the release of entrapped calcein, a fluorescent marker...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(96)79622-4

    authors: Benachir T,Lafleur M

    更新日期:1996-02-01 00:00:00

  • Biophysical evidence that light adaptation in Limulus photoreceptors is due to a negative feedback.

    abstract::The steady-state stimulus-response curve of the Limulus ventral photoreceptor comprises a linear portion followed by a less-than-unity power law dependence, which is maintained over at least 4 decades of intensity. This progressive desensitization corresponds to light adaptation. For flash stimulation of dark-adapted ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(88)83111-4

    authors: Grzywacz NM,Hillman P

    更新日期:1988-03-01 00:00:00

  • The diffusion of oxygen, carbon dioxide, and inert gas in flowing blood.

    abstract::Measurements were made of exchange rates of oxygen, carbon dioxide, and krypton-85 with blood at 37.5 degrees C. Gas transfer took place across a 1 mil silicone rubber membrane. The blood was in a rotating disk boundary layer flow, and the controlling resistance to transfer was the concentration boundary layer. Measur...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(67)86624-4

    authors: Spaeth EE,Friedlander SK

    更新日期:1967-11-01 00:00:00

  • Myosin subfragment-1 attachment to actin. Expected effect on equatorial reflections.

    abstract::The characteristic equatorial X-ray pattern from a relaxed vertebrate skeletal muscle changes when the muscle is activated. In particular, there is a simultaneous decrease in the intensity of the first reflection (I10) and increase in the intensity of the second (I11). This observed change is almost reciprocal. When c...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(78)85510-6

    authors: Lymn RW

    更新日期:1978-01-01 00:00:00

  • Backbone structure of the amantadine-blocked trans-membrane domain M2 proton channel from Influenza A virus.

    abstract::Amantadine is known to block the M2 proton channel of the Influenza A virus. Here, we present a structure of the M2 trans-membrane domain blocked with amantadine, built using orientational constraints obtained from solid-state NMR polarization-inversion-spin-exchange-at-the-magic-angle experiments. The data indicates ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.106.090183

    authors: Hu J,Asbury T,Achuthan S,Li C,Bertram R,Quine JR,Fu R,Cross TA

    更新日期:2007-06-15 00:00:00

  • Electrostatic coupling between retinal isomerization and the ionization state of Glu-204: a general mechanism for proton release in bacteriorhodopsin.

    abstract::The pKa values of ionizable groups that lie between the active site region of bacteriorhodopsin (bR) and the extracellular surface of the protein are reported. Glu-204 is found to have an elevated pKa in the resting state of bR, suggesting that it corresponds to the proton-releasing group in bR. Its elevated pKa is pr...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(96)79320-7

    authors: Sampogna RV,Honig B

    更新日期:1996-09-01 00:00:00

  • Modulation of titin-based stiffness by disulfide bonding in the cardiac titin N2-B unique sequence.

    abstract::The giant protein titin is responsible for the elasticity of nonactivated muscle sarcomeres. Titin-based passive stiffness in myocardium is modulated by titin-isoform switching and protein-kinase (PK)A- or PKG-dependent titin phosphorylation. Additional modulatory effects on titin stiffness may arise from disulfide bo...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2009.05.037

    authors: Grützner A,Garcia-Manyes S,Kötter S,Badilla CL,Fernandez JM,Linke WA

    更新日期:2009-08-05 00:00:00

  • The solution structure of the four-way DNA junction at low-salt conditions: a fluorescence resonance energy transfer analysis.

    abstract::The four-way DNA (Holliday) junction is an important postulated intermediate in the process of genetic recombination. Earlier studies have suggested that the junction exists in two alternative conformations, depending upon the salt concentration present. At high salt concentrations the junction folds into a stacked X ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(94)80765-9

    authors: Clegg RM,Murchie AI,Lilley DM

    更新日期:1994-01-01 00:00:00

  • Structural Thermodynamics of myr-Src(2-19) Binding to Phospholipid Membranes.

    abstract::Many proteins are anchored to lipid bilayer membranes through a combination of hydrophobic and electrostatic interactions. In the case of the membrane-bound nonreceptor tyrosine kinase Src from Rous sarcoma virus, these interactions are mediated by an N-terminal myristoyl chain and an adjacent cluster of six basic ami...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2015.06.051

    authors: Scheidt HA,Klingler J,Huster D,Keller S

    更新日期:2015-08-04 00:00:00

  • Transmembrane helix stability: the effect of helix-helix interactions studied by Fourier transform infrared spectroscopy.

    abstract::We have measured, using infrared spectroscopy, the hydrogen/deuterium exchange rates of the amide protons in the photosynthetic antenna of Rhodospirillum rubrum. These measurements were made not only on the intact protein in detergent solution but also on two dissociated forms (B820 and B777). We have, on the basis of...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(98)74022-6

    authors: Sturgis J,Robert B,Goormaghtigh E

    更新日期:1998-02-01 00:00:00

  • Molecular dynamics simulation of dipalmitoylphosphatidylcholine membrane with cholesterol sulfate.

    abstract::Using the molecular dynamics simulation technique, we studied the changes occurring in a dipalmitoylphosphatidylcholine (DPPC):cholesterol (CH) membrane at 50 mol% sterol when cholesterol is replaced with cholesterol sulfate (CS). Our simulations were performed at constant pressure and temperature on a nanosecond time...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(00)76719-1

    authors: Smondyrev AM,Berkowitz ML

    更新日期:2000-04-01 00:00:00

  • How to make tubular crystals by reconstitution of detergent-solubilized Ca2(+)-ATPase.

    abstract::In an attempt to better define the parameters governing reconstitution and two-dimensional crystallization of membrane proteins, we have studied Ca2(+)-ATPase from rabbit sarcoplasmic reticulum. This ion pump forms vanadate-induced crystals in its native membrane and has previously been reconstituted at high lipid-to-...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(97)78898-2

    authors: Young HS,Rigaud JL,Lacapère JJ,Reddy LG,Stokes DL

    更新日期:1997-06-01 00:00:00

  • Peptides derived from apoptotic Bax and Bid reproduce the poration activity of the parent full-length proteins.

    abstract::Bax and Bid are proapoptotic proteins of the Bcl-2 family that regulate the release of apoptogenic factors from mitochondria. Although they localize constitutively in the cytoplasm, their apoptotic function is exerted at the mitochondrial outer membrane, and is related to their ability to form transbilayer pores. Here...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.104.058008

    authors: García-Sáez AJ,Coraiola M,Dalla Serra M,Mingarro I,Menestrina G,Salgado J

    更新日期:2005-06-01 00:00:00

  • Structural variation in human apolipoprotein E3 and E4: secondary structure, tertiary structure, and size distribution.

    abstract::Human apolipoprotein E (apoE) is a 299-amino-acid protein with a molecular weight of 34 kDa. The difference between the apoE3 and apoE4 isoforms is a single residue substitution involving a Cys-Arg replacement at residue 112. ApoE4 is positively associated with atherosclerosis and late-onset and sporadic Alzheimer's d...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.104.046813

    authors: Chou CY,Lin YL,Huang YC,Sheu SY,Lin TH,Tsay HJ,Chang GG,Shiao MS

    更新日期:2005-01-01 00:00:00

  • Passive mechanical behavior of human neutrophils: effect of cytochalasin B.

    abstract::Actin is a ubiquitous protein in eukaryotic cells. It plays a major role in cell motility and in the maintenance and control of cell shape. In this article, we intend to address the contribution of actin to the passive mechanical properties of human neutrophils. As a framework for assessing this contribution, the neut...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(94)81012-4

    authors: Tsai MA,Frank RS,Waugh RE

    更新日期:1994-06-01 00:00:00

  • Toward prediction of the local onset of alternans in the heart.

    abstract::A beat-to-beat variation in the cardiac action potential duration is a phenomenon known as alternans. Alternans has been linked to ventricular fibrillation, and thus the ability to predict the onset of alternans could be clinically beneficial. Theoretically, it has been proposed that the slope of a restitution curve, ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2011.01.009

    authors: Cram AR,Rao HM,Tolkacheva EG

    更新日期:2011-02-16 00:00:00

  • Effect of Phosphorylation on a Human-like Osteopontin Peptide.

    abstract::The last decade established that the dynamic properties of the phosphoproteome are central to function and its modulation. The temporal dimension of phosphorylation effects remains nonetheless poorly understood, particularly for intrinsically disordered proteins. Osteopontin, selected for this study due to its key rol...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2017.03.005

    authors: Lenton S,Grimaldo M,Roosen-Runge F,Schreiber F,Nylander T,Clegg R,Holt C,Härtlein M,García Sakai V,Seydel T,Marujo Teixeira SC

    更新日期:2017-04-25 00:00:00

  • Kinetic model for the interaction of myosin subfragment 1 with regulated actin.

    abstract::A one-dimensional kinetic Ising model is developed to describe the binding of myosin subfragment 1 (SF-1) to regulated actin. The model allows for cooperative interactions between individual actin sites with bound SF-1 ligands rather than assuming that groups of actin monomer sites change their state in a cooperative ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(83)84286-6

    authors: Balazs AC,Epstein IR

    更新日期:1983-11-01 00:00:00

  • The conduction of protons in different stereoisomers of dioxolane-linked gramicidin A channels.

    abstract::Two different stereoisomers of the dioxolane-linked gramicidin A (gA) channels were individually synthesized (the SS and RR dimers;. Science. 244:813-817). The structural differences between these dimers arise from different chiralities within the dioxolane linker. The SS dimer mimics the helicity and the inter- and i...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(99)77084-0

    authors: Quigley EP,Quigley P,Crumrine DS,Cukierman S

    更新日期:1999-11-01 00:00:00

  • Influence of a natural and a synthetic inhibitor of factor XIIIa on fibrin clot rheology.

    abstract::We investigated the origins of greater clot rigidity associated with FXIIIa-dependent cross-linking. Fibrin clots were examined in which cross-linking was controlled through the use of two inhibitors: a highly specific active-center-directed synthetic inhibitor of FXIIIa, 1,3-dimethyl-4,5-diphenyl-2[2(oxopropyl)thio]i...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(99)77114-6

    authors: Ryan EA,Mockros LF,Stern AM,Lorand L

    更新日期:1999-11-01 00:00:00

  • Unfolding of acrylodan-labeled human serum albumin probed by steady-state and time-resolved fluorescence methods.

    abstract::Steady-state and time-resolved fluorescence spectroscopy was used to follow the local and global changes in structure and dynamics during chemical and thermal denaturation of unlabeled human serum albumin (HSA) and HSA with an acrylodan moiety bound to Cys34. Acrylodan fluorescence was monitored to obtain information ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(98)77598-8

    authors: Flora K,Brennan JD,Baker GA,Doody MA,Bright FV

    更新日期:1998-08-01 00:00:00

  • Effect of noise on DNA sequencing via transverse electronic transport.

    abstract::Previous theoretical studies have shown that measuring the transverse current across DNA strands while they translocate through a nanopore or channel may provide a statistically distinguishable signature of the DNA bases, and may thus allow for rapid DNA sequencing. However, fluctuations of the environment, such as io...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2009.06.055

    authors: Krems M,Zwolak M,Pershin YV,Di Ventra M

    更新日期:2009-10-07 00:00:00

  • A systematic methodology for defining coarse-grained sites in large biomolecules.

    abstract::Coarse-grained (CG) models of biomolecules have recently attracted considerable interest because they enable the simulation of complex biological systems on length-scales and timescales that are inaccessible for atomistic molecular dynamics simulation. A CG model is defined by a map that transforms an atomically detai...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.108.139626

    authors: Zhang Z,Lu L,Noid WG,Krishna V,Pfaendtner J,Voth GA

    更新日期:2008-12-01 00:00:00

  • Proteins in membrane mimetic systems. Insertion of myelin basic protein into microemulsion droplets.

    abstract::The insertion of myelin basic protein into microemulsion droplets of sodium bis (2-ethylhexyl) sulfosuccinate (AOT) has been studied by quasi-elastic light scattering. Measurements were made at both low and high molar ratios of water to surfactant, as a function of protein occupancy. The hydrodynamic radii of filled a...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(85)83851-0

    authors: Chatenay D,Urbach W,Cazabat AM,Vacher M,Waks M

    更新日期:1985-12-01 00:00:00

  • Bilayer conformation of fusion peptide of influenza virus hemagglutinin: a molecular dynamics simulation study.

    abstract::Unraveling the conformation of membrane-bound viral fusion peptides is essential for understanding how those peptides destabilize the bilayer topology of lipids that is important for virus-cell membrane fusion. Here, molecular dynamics (MD) simulations were performed to investigate the conformation of the 20 amino aci...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.103.024562

    authors: Huang Q,Chen CL,Herrmann A

    更新日期:2004-07-01 00:00:00

  • Calcium transients in cerebellar granule cell presynaptic terminals.

    abstract::Calcium ions act presynaptically to modulate synaptic strength and to trigger neurotransmitter release. Here we detect stimulus-evoked changes in residual free calcium ([Ca2+]i) in rat cerebellar granule cell presynaptic terminals. Granule cell axons, known as parallel fibers, and their associated boutons, were labele...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(95)80398-X

    authors: Regehr WG,Atluri PP

    更新日期:1995-05-01 00:00:00