Accessibility of the CLC-0 pore to charged methanethiosulfonate reagents.

Abstract:

:Using the substituted-cysteine-accessibility method, we previously showed that a cysteine residue introduced to the Y512 position of CLC-0 was more rapidly modified by a negatively charged methanethiosulfonate (MTS) reagent, 2-sulfonatoethyl MTS (MTSES), than by the positively charged 2-(trimethylammonium)ethyl MTS (MTSET). This result suggests that a positive intrinsic pore potential attracts the negatively charged MTS molecule. In this study, we further test this hypothesis of a positive pore potential in CLC-0 and find that the preference for the negatively charged MTS is diminished significantly in modifying the substituted cysteine at a deeper pore position, E166. To examine this conundrum, we study the rates of MTS inhibitions of the E166C current and those of the control mutant current from E166A. The results suggest that the inhibition of E166C by intracellularly applied MTS reagents is tainted by the modification of an endogenous cysteine, C229, located at the channel's dimer interface. After this endogenous cysteine is mutated, CLC-0 resumes its preference for selecting MTSES in modifying E166C, reconfirming the idea that the pore of CLC-0 is indeed built with a positive intrinsic potential. These experiments also reveal that MTS modification of C229 can inhibit the current of CLC-0 depending on the amino acid placed at position 166.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Zhang XD,Yu WP,Chen TY

doi

10.1016/j.bpj.2009.09.066

subject

Has Abstract

pub_date

2010-02-03 00:00:00

pages

377-85

issue

3

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(09)01632-4

journal_volume

98

pub_type

杂志文章
  • Link between the enzymatic kinetics and mechanical behavior in an actomyosin motor.

    abstract::We have attempted to link the solution actomyosin ATPase with the mechanical properties of in vitro actin filament sliding over heavy meromyosin. To accomplish this we perturbed the system by altering the substrate with various NTPs and divalent cations, and by altering ionic strength. A wide variety of enzymatic and ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(01)76022-5

    authors: Amitani I,Sakamoto T,Ando T

    更新日期:2001-01-01 00:00:00

  • A molecular dynamics study of the ligand release path in yeast cytosine deaminase.

    abstract::Yeast cytosine deaminase, a zinc metalloenzyme, catalyzes the deamination of cytosine to uracil. Experimental and computational evidence indicates that the rate-limiting step is product release, instead of the chemical reaction step. In this work, we use molecular dynamics to suggest ligand exit paths. Simulation at 3...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.106.098921

    authors: Yao L,Yan H,Cukier RI

    更新日期:2007-04-01 00:00:00

  • The length dependence of translational diffusion, free solution electrophoretic mobility, and electrophoretic tether force of rigid rod-like model duplex DNA.

    abstract::In this work, boundary element modeling is used to study the transport of highly charged rod-like model polyions of various length under a variety of different aqueous salt conditions. Transport properties considered include free solution electrophoretic mobility, translational diffusion, and the components of the "te...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(01)75900-0

    authors: Allison S,Chen C,Stigter D

    更新日期:2001-11-01 00:00:00

  • Selective detection of the rotational dynamics of the protein-associated lipid hydrocarbon chains in sarcoplasmic reticulum membranes.

    abstract::We have developed a saturation transfer EPR (ST-EPR) method to measure selectively the rotational dynamics of those lipids that are motionally restricted by integral membrane proteins and have applied this methodology to measure lipid-protein interactions in native sarcoplasmic reticulum (SR) membranes. This analysis ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(89)82721-3

    authors: Squier TC,Thomas DD

    更新日期:1989-10-01 00:00:00

  • Structure and dynamics of Escherichia coli chemosensory receptors. Engineered sulfhydryl studies.

    abstract::Cysteine residues introduced by site-directed mutagenesis have been used to probe the conformation and dynamics of two receptors in the E. coli chemotaxis pathway. (a) Thermal motions of the polypeptide backbone were investigated in the periplasmic D-galactose and D-glucose receptor, a globular protein of known struct...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(92)81806-4

    authors: Careaga CL,Falke JJ

    更新日期:1992-04-01 00:00:00

  • Structural rearrangements in the active site of smooth-muscle myosin.

    abstract::Structural rearrangements of the myosin upper-50 kD subdomain are thought to play a key role in coordinating actin binding with nucleotide hydrolysis during the myosin ATPase cycle. Such rearrangements could open and close the active site in opposition to the actin-binding cleft, helping explain the opposing affinitie...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.105.059840

    authors: Robertson CI,Gaffney DP 2nd,Chrin LR,Berger CL

    更新日期:2005-09-01 00:00:00

  • Impulse responses of automaticity in the Purkinje fiber.

    abstract::We examined the effects of brief current pulses on the pacemaker oscillations of the Purkinje fiber using the model of McAllister , Noble, and Tsien (1975. J. Physiol. [Lond.]. 251:1-57). This model was used to construct phase-response curves for brief electric stimuli to find "black holes," where rhythmic activity of...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(84)84228-9

    authors: Chay TR,Lee YS

    更新日期:1984-04-01 00:00:00

  • Measurement of lipid nanodomain (raft) formation and size in sphingomyelin/POPC/cholesterol vesicles shows TX-100 and transmembrane helices increase domain size by coalescing preexisting nanodomains but do not induce domain formation.

    abstract::Mixtures of unsaturated lipids, sphingolipids, and cholesterol form coexisting liquid-disordered and sphingolipid and cholesterol-rich liquid-ordered (Lo) phases in water. The detergent Triton X-100 does not readily solubilize Lo domains, but does solubilize liquid-disordered domains, and is commonly used to prepare d...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2011.08.059

    authors: Pathak P,London E

    更新日期:2011-11-16 00:00:00

  • Molecular dynamics simulations reveal multiple pathways of ligand dissociation from thyroid hormone receptors.

    abstract::Nuclear receptor (NR) ligands occupy a pocket that lies within the core of the NR ligand-binding domain (LBD), and most NR LBDs lack obvious entry/exit routes upon the protein surface. Thus, significant NR conformational rearrangements must accompany ligand binding and release. The precise nature of these processes, h...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.105.063818

    authors: Martínez L,Sonoda MT,Webb P,Baxter JD,Skaf MS,Polikarpov I

    更新日期:2005-09-01 00:00:00

  • Gating of cyclic nucleotide-gated (CNGA1) channels by cGMP jumps and depolarizing voltage steps.

    abstract::We expressed rod-type homotetrameric cyclic nucleotide-gated (CNGA1) channels in Xenopus oocytes and studied activation by photolysis-induced jumps of the 3',5'-cyclic guanosine monophosphate (cGMP) concentration and by voltage steps. cGMP jumps to increasing concentrations up to the EC50 value of 46.5 microM decelera...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.105.078667

    authors: Nache V,Kusch J,Hagen V,Benndorf K

    更新日期:2006-05-01 00:00:00

  • Toward the understanding of MNEI sweetness from hydration map surfaces.

    abstract::The binding mechanism of sweet proteins to their receptor, a G-protein-coupled receptor, is not supported by direct structural information. In principle, the key groups responsible for biological activity (glucophores) can be localized on a small structural unit (sweet finger) or spread on a larger surface area. A rec...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.105.073171

    authors: De Simone A,Spadaccini R,Temussi PA,Fraternali F

    更新日期:2006-05-01 00:00:00

  • Birefringence of protein solutions and biological systems. II. Studies on TMV, tropocollagen, and paramyosin.

    abstract::The intrinsic birefringences of TMV, tropocollagen, and paramyosin were calculated from flow birefringence measurements using the theory of Peterlin and Stuart. The values are -0.029, -0.029, and -0.030, respectively. The intrinsic birefringences of TMV and tropocollagen were measured as a function of the refractive i...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/s0006-3495(63)86810-1

    authors: TAYLOR EW,CRAMER W

    更新日期:1963-03-01 00:00:00

  • Molecular dynamics study of the energetic, mechanistic, and structural implications of a closed phosphate tube in ncd.

    abstract::The switch 1 region of myosin forms a lid over the nucleotide phosphates as part of a structure known as the phosphate-tube. The homologous region in kinesin-family motors is more open, not interacting with the nucleotide. We used molecular dynamics (MD) simulations to examine a possible displacement of switch 1 of th...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(01)76092-4

    authors: Minehardt TJ,Cooke R,Pate E,Kollman PA

    更新日期:2001-03-01 00:00:00

  • Inhibition of pulmonary surfactant adsorption by serum and the mechanisms of reversal by hydrophilic polymers: theory.

    abstract::A theory based on the Smolukowski analysis of colloid stability shows that the presence of charged, surface-active serum proteins at the alveolar air-liquid interface can severely reduce or eliminate the adsorption of lung surfactant from the subphase to the interface, consistent with the observations reported in the ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.105.062646

    authors: Zasadzinski JA,Alig TF,Alonso C,Bernardino de la Serna J,Perez-Gil J,Taeusch HW

    更新日期:2005-09-01 00:00:00

  • An estimated shape function for drift in a platelet-transport model.

    abstract::Prior work has shown that concentration profiles of platelets in flowing whole blood and of platelet-sized beads in flowing blood suspensions can include near-wall excesses. A model to describe this phenomenon was built about a single-component convective diffusion equation. To incorporate redistribution to preferred ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(94)80595-8

    authors: Yeh C,Calvez AC,Eckstein EC

    更新日期:1994-09-01 00:00:00

  • Water Distribution within Wild-Type NRas Protein and Q61 Mutants during Unrestrained QM/MM Dynamics.

    abstract::Point mutations in p21ras are associated with ∼30% of human tumors by disrupting its GTP hydrolysis cycle, which is critical to its molecular switch function in cellular signaling pathways. In this work, we investigate the impact of Gln 61 substitutions in the structure of the p21N-ras active site and particularly foc...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2018.07.042

    authors: Tichauer RH,Favre G,Cabantous S,Landa G,Hemeryck A,Brut M

    更新日期:2018-10-16 00:00:00

  • Ligand-receptor kinetics measured by total internal reflection with fluorescence correlation spectroscopy.

    abstract::Total internal reflection excitation used in combination with fluorescence correlation spectroscopy (TIR-FCS) is a method for characterizing the dynamic behavior and absolute concentrations of fluorescent molecules near or at the interface of a planar substrate and a solution. In this work, we demonstrate for the firs...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(03)74748-1

    authors: Lieto AM,Cush RC,Thompson NL

    更新日期:2003-11-01 00:00:00

  • Nanoparticles as fluorescence labels: is size all that matters?

    abstract::Fluorescent labels are often used in bioassays as a means to detect and characterize ligand-receptor binding. This is due in part to the inherently high sensitivity of fluorescence-based technology and the relative accessibility of the technique. There is often little concern raised as to whether or not the fluorescen...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.107.127688

    authors: Swift JL,Cramb DT

    更新日期:2008-07-01 00:00:00

  • Modeling the step of endosomal escape during cell infection by a nonenveloped virus.

    abstract::Widely disparate viruses enter the host cell through an endocytic pathway and travel the cytoplasm inside an endosome. For the viral genetic material to be delivered into the cytoplasm, these viruses have to escape the endosomal compartment, an event triggered by the conformational changes of viral endosomolytic prote...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2011.12.037

    authors: Lagache T,Danos O,Holcman D

    更新日期:2012-03-07 00:00:00

  • Procedure for testing kinetic models of the photocycle of bacteriorhodopsin.

    abstract::Given some simple kinetic models of the photocycle of bacteriorhodopsin (bR) and data taken at many wavelengths and under conditions that avoid photoselection and steady-state cycling complications, it is shown how to extract the apparent rate constants and the spectra of the intermediates. Special consideration was g...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(82)84543-8

    authors: Nagle JF,Parodi LA,Lozier RH

    更新日期:1982-05-01 00:00:00

  • An analytic solution to the Monod-Wyman-Changeux model and all parameters in this model.

    abstract::Starting from the Monod-Wyman-Changeux (MWC) model (Monod, J., J. Wyman, and J. P. Changeux. 1965. J. Mol. Biol. 12:88-118), we obtain an analytical expression for the slope of the Hill plot at any ligand concentration. Furthermore, we derive an equation satisfied by the ligand concentration at the position of maximum...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(89)82802-4

    authors: Zhou G,Ho PS,van Holde KE

    更新日期:1989-02-01 00:00:00

  • In situ analysis of spatial relationships between proteins of the nuclear pore complex.

    abstract::Macromolecular transport between the nucleus and cytoplasm occurs through the nuclear pore complexes (NPCs). The NPC in the budding yeast Saccharomyces cerevisiae is a 60-MDa structure embedded in the nuclear envelope and composed of ~30 proteins, termed nucleoporins or nups. Here we present a large-scale analysis of ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(02)75363-0

    authors: Damelin M,Silver PA

    更新日期:2002-12-01 00:00:00

  • Receptor displacement in the cell membrane by hydrodynamic force amplification through nanoparticles.

    abstract::We introduce an intrinsically multiplexed and easy to implement method to apply an external force to a biomolecule and thus probe its interaction with a second biomolecule or, more generally, its environment (for example, the cell membrane). We take advantage of the hydrodynamic interaction with a controlled fluid flo...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2013.05.045

    authors: Türkcan S,Richly MU,Bouzigues CI,Allain JM,Alexandrou A

    更新日期:2013-07-02 00:00:00

  • Behavior of supercoiled DNA.

    abstract::We study DNA supercoiling in a quantitative fashion by micromanipulating single linear DNA molecules with a magnetic field gradient. By anchoring one end of the DNA to multiple sites on a magnetic bead and the other end to multiple sites on a glass surface, we were able to exert torsional control on the DNA. A rotatin...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(98)77908-1

    authors: Strick TR,Allemand JF,Bensimon D,Croquette V

    更新日期:1998-04-01 00:00:00

  • Mechanosensitivity Occurs along the Adhesome's Force Train and Affects Traction Stress.

    abstract::Herein, we consider the process of force development along the adhesome within cell focal adhesions. Our model adhesome consists of the actin cytoskeleton-vinculin-talin-integrin-ligand-extracellular matrix-substrate force train. We specifically consider the effects of substrate stiffness on the force levels expected ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2019.08.039

    authors: Asaro RJ,Lin K,Zhu Q

    更新日期:2019-11-05 00:00:00

  • Modifications of sodium channel gating in Myxicola giant axons by deuterium oxide, temperature, and internal cations.

    abstract::In dialyzed Myxicola axons substitution of heavy water (D2O) externally and internally slows both sodium and potassium kinetics and decreases the maximum conductances. Furthermore, this effect is strongly temperature dependent, the magnitude of the slowing produced by D2O substitution decreasing with increasing temper...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(79)85211-X

    authors: Schauf CL,Bullock JO

    更新日期:1979-08-01 00:00:00

  • Light-induced voltage changes associated with electron and proton transfer in photosystem II core complexes reconstituted in phospholipid monolayers.

    abstract::We have measured light-induced voltage changes (electrogenic events) in photosystem II (PSII) core complexes oriented in phospholipid monolayers. These events are compared to those measured in the functionally and structurally closely related reaction centers from the photosynthetic bacterium Rhodobacter sphaeroides. ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(94)81001-X

    authors: Höök F,Brzezinski P

    更新日期:1994-06-01 00:00:00

  • Reorganization of lipid domain structure in membranes by a transmembrane peptide: an ESR spin label study on the effect of the Escherichia coli outer membrane protein A signal peptide on the fluid lipid domain connectivity in binary mixtures of dimyristoy

    abstract::The effect of a transmembrane peptide on the domain structure of a two-component, two-phase lipid bilayer composed of dimyristoyl phosphatidylcholine (DMPC) and distearoyl phosphatidylcholine (DSPC) was examined by spin label electron spin resonance (ESR) spectroscopy. The peptide, pOmpA, is the hydrophobic, 25-residu...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(94)80989-0

    authors: Sankaram MB,Marsh D,Gierasch LM,Thompson TE

    更新日期:1994-06-01 00:00:00

  • Inversion of proton translocation in bacteriorhodopsin mutants D85N, D85T, and D85,96N.

    abstract::Proton translocation activity of bacteriorhodopsin mutants lacking the proton acceptor Asp-85 was investigated using the black lipid membrane technique. Mutants D85N, D85T, and D85,96N were constructed and homologously expressed in Halobacterium salinarium to yield a membrane fraction with a buoyant density of 1.18 g/...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(94)80642-3

    authors: Tittor J,Schweiger U,Oesterhelt D,Bamberg E

    更新日期:1994-10-01 00:00:00

  • An information-based approach to change-point analysis with applications to biophysics and cell biology.

    abstract::This article describes the application of a change-point algorithm to the analysis of stochastic signals in biological systems whose underlying state dynamics consist of transitions between discrete states. Applications of this analysis include molecular-motor stepping, fluorophore bleaching, electrophysiology, partic...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2015.05.038

    authors: Wiggins PA

    更新日期:2015-07-21 00:00:00