Fluorescence study of the divalent cation-transport mechanism of ionophore A23187 in phospholipid membranes.

Abstract:

:The mechanism for transport of divalent cations across phospholipid bilayers by the ionophore A23187 was investigated. The intrinsic fluorescence of the ionophore was used in equilibrium and rapid-mixing experiments as an indicator of ionophore environment and complexation with divalent cations. The neutral (protonated) form of the ionophore binds strongly to the membrane, with a high quantum yield relative to that in the aqueous phase. The negatively charged form of the ionophore binds somewhat less strongly, with a lower quantum yield, and does not move across the membrane. Complexation of the negatively charged form with divalent cations was measured by the decrease in fluorescence. An apparent rate constant (kapp) for transport of the ionophore across the membrane was determined from the rate of fluorescence changes observed in stopped-flow rapid kinetic experiments. The variation of kapp was studied as a function of pH, temperature, ionophore concentration, membrane lipid composition, and divalent cation concentration and type. Analysis and comparison with equilibrium constants for protonation and complexation show that A23187 and its metal:ionophore complexes bind near the membrane-water interface in the lipid polar-head region. The interfacial reactions occur rapidly, compared with the transmembrane reactions, and are thus in equilibrium during transport. The transport cycle can be described as follows: a 1:1 complex is formed between the membrane bound A23187-(Am-) and the aqueous divalent cation with dissociation constant K1 approximately 4.6 x 10(-4) M. This is in equilibrium with a 1:2 (metal:ionophore) complex (K2 approximately 3.0 x 10(-4) [ionophore/lipid]) that is responsible for transporting the divalent cations across the membrane. The rate constant for translocation of the 1:2 complex is 0.1-0.3 s-1. Dissociation of the complex of the trans side and protonation occur rapidly. The rate constant for translocation of H+ . A23187- is 28 s-1. A theory is presented that is capable of reproducing the kinetic data at any calcium concentration. The cation specificity for ionophore complex transport (kapp), determined at low ionophore concentration for a series of divalent cations, was found to be proportional to the equilibrium constant for 1:1 complexation. The order of ion specificity for these processes was found to be Ca2+ greater than Mg2+ greater Sr2+ greater than Ba2+. Interactions with Na+ were not observed. Maximal values of kapp were observed for vesicles prepared from pure dimyristoyl phosphatidylcholine. Inclusion of phosphatidyl ethanolamine, phosphatidic acid, or dipalmatoyl phosphatidylcholine resulted in lower values of kapp. Calcium transport by A23187 is compared with that of X537A, and it is shown that the former is 67-fold faster. The difference in rates is due to differences in the ability of each ionophore to form a 1:2 complex from a 1:1 complex.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Kolber MA,Haynes DH

doi

10.1016/S0006-3495(81)84738-8

subject

Has Abstract

pub_date

1981-11-01 00:00:00

pages

369-91

issue

2

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(81)84738-8

journal_volume

36

pub_type

杂志文章
  • Characterization of permeability and morphological perturbations induced by nisin on phosphatidylcholine membranes.

    abstract::Nisin is an antimicrobial peptide used as food preservative. To gain some insights into the hypothesis that its bactericidal activity is due to the perturbation of the lipid fraction of the bacterial plasmic membrane, we have investigated the effect of nisin on model phosphatidylcholine (PC) membranes. We show that ni...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(99)76936-5

    authors: El Jastimi R,Edwards K,Lafleur M

    更新日期:1999-08-01 00:00:00

  • Optical and physical properties of wet-spun films of Na-hyaluronate. Evidence of a phase transition.

    abstract::The refractive indices, water content, and volume of wet-spun films of Na-hyaluronate have been measured as a function of relative humidity (rh). These data are used with the Lorentz-Lorenz formula to determine the optical polarizabilities of Na-hyaluronate parallel and perpendicular to the helical axis. The analysis ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(92)81604-1

    authors: Flowers MR,Marlow RL,Lee SA,Lavalle N,Rupprecht A

    更新日期:1992-08-01 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

  • Osmotic pressure and packaging structure of caged DNA.

    abstract::We present a theoretical model for aqueous solutions of double-stranded (ds) DNA with explicit consideration of electrostatic interactions, excluded-volume effects, van der Waals attractions, and salt ions. With reasonable parameters estimated from the DNA structure and experimental data for electrolytes, we are able ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.107.112508

    authors: Li Z,Wu J,Wang ZG

    更新日期:2008-02-01 00:00:00

  • Regulation of calcium channel activity by lipid domain formation in planar lipid bilayers.

    abstract::The sarcoplasmic reticulum channel (ryanodine receptor) from cardiac myocytes was reconstituted into planar lipid bilayers consisting of 1-palmitoyl-2-oleoyl-phosphatidylethanolamine (POPE) and 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) in varying ratios. The channel activity parameters, i.e., open probability an...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(03)74532-9

    authors: Cannon B,Hermansson M,Györke S,Somerharju P,Virtanen JA,Cheng KH

    更新日期:2003-08-01 00:00:00

  • Membrane tension, myosin force, and actin turnover maintain actin treadmill in the nerve growth cone.

    abstract::A growth cone is a motile structure at the tips of axons that is driven by the actin network and guides axon extension. Low actin adhesion to the substrate creates a stationary actin treadmill that allows leading-edge protrusion when adhesion increases in response to guidance cues. We use experimental measurements in ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2012.03.003

    authors: Craig EM,Van Goor D,Forscher P,Mogilner A

    更新日期:2012-04-04 00:00:00

  • Mechanism of pH-triggered collapse of phosphatidylethanolamine liposomes stabilized by an ortho ester polyethyleneglycol lipid.

    abstract::The mechanism of pH-triggered destabilization of liposomes composed of a polyethyleneglycol-orthoester-distearoylglycerol lipid (POD) and phosphatidyl ethanolamine (PE) has been studied using an ANTS/DPX leakage and a lipid-mixing assay. We developed a kinetic model that relates POD hydrolysis to liposome collapse. Th...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/s0006-3495(03)74986-8

    authors: Guo X,MacKay JA,Szoka FC Jr

    更新日期:2003-03-01 00:00:00

  • Calorimetric studies of bovine rod outer segment disk membranes support a monomeric unit for both rhodopsin and opsin.

    abstract::The photoreceptor rhodopsin is a G-protein coupled receptor that has recently been proposed to exist as a dimer or higher order oligomer, in contrast to the previously described monomer, in retinal rod outer segment disk membranes. Rhodopsin exhibits considerably greater thermal stability than opsin (the bleached form...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.108.128868

    authors: Edrington TC 5th,Bennett M,Albert AD

    更新日期:2008-09-15 00:00:00

  • The structural basis of cholesterol accessibility in membranes.

    abstract::Although the majority of free cellular cholesterol is present in the plasma membrane, cholesterol homeostasis is principally regulated through sterol-sensing proteins that reside in the cholesterol-poor endoplasmic reticulum (ER). In response to acute cholesterol loading or depletion, there is rapid equilibration betw...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2013.08.042

    authors: Olsen BN,Bielska AA,Lee T,Daily MD,Covey DF,Schlesinger PH,Baker NA,Ory DS

    更新日期:2013-10-15 00:00:00

  • Noninvasive imaging of the distribution in oxygen in tissue in vivo using near-infrared phosphors.

    abstract::A newly developed water-soluble phosphor suitable for measuring oxygen pressure in the blood (Green 2W) was used for noninvasive, in vivo imaging of oxygen distribution in the vascular systems of mice. Oxygen quenches the phosphorescence of Green 2W, measured in the presence of 2% albumin, according to the Stern-volme...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(96)79764-3

    authors: Vinogradov SA,Lo LW,Jenkins WT,Evans SM,Koch C,Wilson DF

    更新日期:1996-04-01 00:00:00

  • Prediction of solution structures of the Ca2+-bound gamma-carboxyglutamic acid domains of protein S and homolog growth arrest specific protein 6: use of the particle mesh Ewald method.

    abstract::The solution structures of the N-terminal domains of protein S, a plasma vitamin K-dependent glycoprotein, and its homolog growth arrest specific protein 6 (Gas6) were predicted by molecular dynamics computer simulations. The initial structures were based on the x-ray crystallographic structure of the corresponding re...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(97)78215-8

    authors: Perera L,Li L,Darden T,Monroe DM,Pedersen LG

    更新日期:1997-10-01 00:00:00

  • Influence of ionic strength on the actomyosin reaction steps in contracting skeletal muscle fibers.

    abstract::Muscle contraction occurs as the result of actin-myosin interaction, which is mediated by the intermolecular forces exerted at the actin-myosin interface. To obtain information about the nature of these intermolecular forces, we tested the sensitivity of various contractile parameters of skinned skeletal muscle fibers...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(00)76850-0

    authors: Iwamoto H

    更新日期:2000-06-01 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

  • Modeling of Transmembrane Potential in Realistic Multicellular Structures before Electroporation.

    abstract::Many approaches for studying the transmembrane potential (TMP) induced during the treatment of biological cells with pulsed electric fields have been reported. From the simple analytical models to more complex numerical models requiring significant computational resources, a gamut of methods have been used to recapitu...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2016.10.005

    authors: Murovec T,Sweeney DC,Latouche E,Davalos RV,Brosseau C

    更新日期:2016-11-15 00:00:00

  • Cell-cell interactions mediate the response of vascular smooth muscle cells to substrate stiffness.

    abstract::The vessel wall experiences progressive stiffening with age and the development of cardiovascular disease, which alters the micromechanical environment experienced by resident vascular smooth muscle cells (VSMCs). In vitro studies have shown that VSMCs are sensitive to substrate stiffness, but the exact molecular mech...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2011.06.051

    authors: Sazonova OV,Lee KL,Isenberg BC,Rich CB,Nugent MA,Wong JY

    更新日期:2011-08-03 00:00:00

  • Mathematical analysis and quantification of fluorescent proteins as transcriptional reporters.

    abstract::Fluorescent proteins are often used as reporters of transcriptional activity. Here we present a mathematical characterization of a novel fluorescent reporter that was recently engineered to have a short half-life (approximately 12 min). The advantage of this destabilized protein is that it can track the transient tran...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.107.122200

    authors: Wang X,Errede B,Elston TC

    更新日期:2008-03-15 00:00:00

  • A model of effective diffusion and tortuosity in the extracellular space of the brain.

    abstract::Tortuosity of the extracellular space describes hindrance posed to the diffusion process by a geometrically complex medium in comparison to an environment free of any obstacles. Calculating tortuosity in biologically relevant geometries is difficult. Yet this parameter has proved very important for many processes in t...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.103.039495

    authors: Hrabe J,Hrabetová S,Segeth K

    更新日期:2004-09-01 00:00:00

  • Fibrin Networks Support Recurring Mechanical Loads by Adapting their Structure across Multiple Scales.

    abstract::Tissues and cells sustain recurring mechanical loads that span a wide range of loading amplitudes and timescales as a consequence of exposure to blood flow, muscle activity, and external impact. Both tissues and cells derive their mechanical strength from fibrous protein scaffolds, which typically have a complex hiera...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2016.06.034

    authors: Kurniawan NA,Vos BE,Biebricher A,Wuite GJ,Peterman EJ,Koenderink GH

    更新日期:2016-09-06 00:00:00

  • Single-Molecule Stretching Shows Glycosylation Sets Tension in the Hyaluronan-Aggrecan Bottlebrush.

    abstract::Large bottlebrush complexes formed from the polysaccharide hyaluronan (HA) and the proteoglycan aggrecan contribute to cartilage compression resistance and are necessary for healthy joint function. A variety of mechanical forces act on these complexes in the cartilage extracellular matrix, motivating the need for a qu...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2020.08.016

    authors: Innes-Gold SN,Berezney JP,Saleh OA

    更新日期:2020-10-06 00:00:00

  • Doubly Spin-Labeled RNA as an EPR Reporter for Studying Multicomponent Supramolecular Assemblies.

    abstract::mRNAs are involved in complicated supramolecular complexes with human 40S and 80S ribosomes responsible for the protein synthesis. In this work, a derivative of nonaribonucleotide pUUCGUAAAA with nitroxide spin labels attached to the 5'-phosphate and to the C8 atom of the adenosine in sixth position (mRNA analog) was ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2015.10.042

    authors: Malygin AA,Graifer DM,Meschaninova MI,Venyaminova AG,Krumkacheva OA,Fedin MV,Karpova GG,Bagryanskaya EG

    更新日期:2015-12-15 00:00:00

  • Charge translocation by the Na+/K+ pump under Na+/Na+ exchange conditions: intracellular Na+ dependence.

    abstract::The effect of intracellular (i) and extracellular (o) Na+ on pre-steady-state transient current associated with Na+/Na+ exchange by the Na+/K+ pump was investigated in the vegetal pole of Xenopus oocytes. Current records in response to 40-ms voltage pulses from -180 to +100 mV in the absence of external Na+ were subtr...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.105.072942

    authors: Holmgren M,Rakowski RF

    更新日期:2006-03-01 00:00:00

  • Light signal transduction pathway from flavin chromophore to the J alpha helix of Arabidopsis phototropin1.

    abstract::In the plant blue-light sensor phototropin, illumination of the chromophoric LOV domains causes activation of the serine/threonine kinase domain. Flavin mononucleotide (FMN) is a chromophore molecule in the two LOV domains (LOV1 and LOV2), but only LOV2 is responsible for kinase activation. Previous studies reported a...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2008.12.3924

    authors: Yamamoto A,Iwata T,Sato Y,Matsuoka D,Tokutomi S,Kandori H

    更新日期:2009-04-08 00:00:00

  • Binding of cations of group IA and IIA to bovine serum amine oxidase: effect on the activity.

    abstract::In this paper, we report on the presence of cation binding areas on bovine serum amine oxidase, where metal ions of the groups IA and IIA, such as Na(+), K(+), Cs(+), Mg(2+), and Ca(2+), bind with various affinities. We found a cation-binding area that influences the enzyme activity if occupied, so that the catalytic ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(02)73983-0

    authors: Di Paolo ML,Scarpa M,Corazza A,Stevanato R,Rigo A

    更新日期:2002-10-01 00:00:00

  • Effects of chain unsaturation on the equation of state for lipid monolayers at the air-water interface.

    abstract::An equation of state for lipid monolayers at the air-water interface is presented, which takes into account the effects of the conformation and the number and position of double bonds of the hydrocarbon chains. The total Hamiltonian of the monolayer is assumed to consist of three terms. Two of them are calculated exac...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(95)79919-2

    authors: Feng SS,MacDonald RC

    更新日期:1995-08-01 00:00:00

  • A quantitative model of thermal stabilization and destabilization of proteins by ligands.

    abstract::Equilibrium binding ligands usually increase protein thermal stability by an amount proportional to the concentration and affinity of the ligand. High-throughput screening for the discovery of drug-like compounds uses an assay based on thermal stabilization. The mathematical description of this stabilization is well d...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.108.134973

    authors: Cimmperman P,Baranauskiene L,Jachimoviciūte S,Jachno J,Torresan J,Michailoviene V,Matuliene J,Sereikaite J,Bumelis V,Matulis D

    更新日期:2008-10-01 00:00:00

  • Simultaneous capacitance and amperometric measurements of exocytosis: a comparison.

    abstract::We measured the exocytotic response induced by flash photolysis of caged compounds in isolated mast cells and chromaffin cells. Vesicle fusion was measured by monitoring the cell membrane capacitance. The release of vesicular contents was followed by amperometry. In response to a GTP gamma S stimulus we found that the...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/S0006-3495(96)79315-3

    authors: Oberhauser AF,Robinson IM,Fernandez JM

    更新日期:1996-08-01 00:00:00

  • Using the bias from flow to elucidate single DNA repair protein sliding and interactions with DNA.

    abstract::We perform single-molecule spatial tracking measurements of a DNA repair protein, the C-terminal domain of Ada (C-Ada) from Escherichia coli, moving on DNA extended by flow. The trajectories of single proteins labeled with a fluorophore are constructed. We analyze single-protein dwell times on DNA for different flow r...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2008.11.021

    authors: Lin Y,Zhao T,Jian X,Farooqui Z,Qu X,He C,Dinner AR,Scherer NF

    更新日期:2009-03-04 00:00:00

  • Microscopic mechanism for experimentally observed anomalous elasticity of DNA in two dimensions.

    abstract::By exploring a recent model in which DNA bending elasticity, described by the wormlike chain model, is coupled to basepair denaturation, we demonstrate that small denaturation bubbles lead to anomalies in the flexibility of DNA at the nanometric scale, when confined in two dimensions (2D), as reported in atomic-force ...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2009.03.035

    authors: Destainville N,Manghi M,Palmeri J

    更新日期:2009-06-03 00:00:00

  • Enhancement of diffusion-controlled reaction rates by surface-induced orientational restriction.

    abstract::We explore the means by which immobilization of a substrate on a surface can increase the rate of a diffusion-controlled enzymatic reaction. A quasichemical approach is developed and compared with Brownian dynamics simulations. We use these methods to show that restricting only the orientation of the enzyme by long-ra...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1529/biophysj.105.072785

    authors: Nag A,Dinner AR

    更新日期:2006-02-01 00:00:00

  • Exploration of Insulin Amyloid Polymorphism Using Raman Spectroscopy and Imaging.

    abstract::We aimed to investigate insulin amyloid fibril polymorphism caused by salt effects and heating temperature and to visualize the structural differences of the polymorphisms in situ using Raman imaging without labeling. The time course monitoring for amyloid formation was carried out in an acidic condition without any s...

    journal_title:Biophysical journal

    pub_type: 杂志文章

    doi:10.1016/j.bpj.2020.04.031

    authors: Ishigaki M,Morimoto K,Chatani E,Ozaki Y

    更新日期:2020-06-16 00:00:00