Mechanochemistry of protein 4.1's spectrin-actin-binding domain: ternary complex interactions, membrane binding, network integration, structural strengthening.
AuthorsDischer DE, Winardi R, Schischmanoff PO, Parra M, Conboy JG, Mohandas N
JournalJ Cell Biol
PubMed ID7642705
Mechanical strength of the red cell membrane is dependent on ternary interactions among the skeletal proteins, spectrin, actin, and protein 4.1. Protein 4.1's spectrin-actin-binding (SAB) domain is specified by an alternatively spliced exon encoding 21 amino acid (aa) and a constitutive exon encoding 59 aa. A series of truncated SAB ... More
Lateral mobility of class I histocompatibility antigens in B lymphoblastoid cell membranes: modulation by cross-linking and effect of cell density.
AuthorsBierer BE, Herrmann SH, Brown CS, Burakoff SJ, Golan DE
JournalJ Cell Biol
PubMed ID3115995
We have studied the lateral mobility of class 1 major histocompatibility complex (MHC) proteins in the membranes of human Epstein-Barr virus-transformed B cells using fluorescence photobleaching recovery. Class I MHC antigens were labeled with either W6/32 monoclonal antibody or its Fab fragment directly conjugated to fluorescein isothiocyanate. The diffusion coefficient ... More
Fabrication of phospholipid bilayer-coated microchannels for on-chip immunoassays.
AuthorsYang T, Jung S, Mao H, Cremer PS
JournalAnal Chem
PubMed ID11199961
'Herein we describe a new class of microfluidic immunoassays based upon solid supported lipid bilayers. Two-dimensionally fluid bilayer material, which can accommodate multivalent binding between surface-bound ligands and aqueous receptors, was coated on the surface of poly(dimethylsiloxane) microchannels. The bilayers contained dinitrophenyl (DNP)-conjugated lipids for binding with bivalent anti-DNP antibodies. ... More
Lateral mobility of lipid analogues and GPI-anchored proteins in supported bilayers determined by fluorescent bead tracking.
AuthorsFein M, Unkeless J, Chuang FY, Sassaroli M, da Costa R, Väänänen H, Eisinger J
JournalJ Membr Biol
PubMed ID8411132
'Lipid analogues and glycosylphosphatidylinositol (GPI)-anchored proteins incorporated in glass-supported phospholipid bilayers (SBL) were coupled to small (30 nm diameter) fluorescent beads whose motion in the liquid phase was tracked by intensified fluorescence video microscopy. Streptavidin (St), covalently attached to the carboxyl modified surface of the polystyrene bead, bound either the ... More
Relationship of lipid rafts to transient confinement zones detected by single particle tracking.
AuthorsDietrich C, Yang B, Fujiwara T, Kusumi A, Jacobson K
JournalBiophys J
PubMed ID11751315
'We examined the physical and chemical characteristics of transient confinement zones (TCZs) that are detected in single particle trajectories of molecules moving within the membrane of C3H 10T1/2 murine fibroblasts and their relationship to "rafts." We studied the lateral movement of different membrane molecules thought to partition to varying degrees ... More
Beta-very low density lipoprotein is sequestered in surface-connected tubules in mouse peritoneal macrophages.
AuthorsMyers JN, Tabas I, Jones NL, Maxfield FR
JournalJ Cell Biol
PubMed ID8253839
'beta-very low density lipoprotein (VLDL) is a large lipoprotein with multiple apoprotein E (apoE) molecules that bind to the LDL receptors on mouse macrophages. Even though they bind to the same receptor, the endocytic processing of beta-VLDL differs from low density lipoprotein (LDL). LDL is rapidly delivered to perinuclear lysosomes ... More
Evidence for phospholipid microdomain formation in liquid crystalline liposomes reconstituted with Escherichia coli lactose permease.
AuthorsLehtonen JY, Kinnunen PK
JournalBiophys J
PubMed ID9138570
'The well-characterized integral membrane protein lactose (lac) permease from Escherichia coli was reconstituted together with trace amounts (molar fraction X = 0.005 of the total phospholipid) of different pyrene-labeled phospholipid analogs into 1-palmitoyl-2-oleoyl-sn-glycero-3-sn-glycero-3-phospho-rac'-glycerol (POPG) liposomes. Effects of lac permease on bilayer lipid dynamics were investigated by measuring the excimer-to-monomer fluorescence ... More
Transient confinement of a glycosylphosphatidylinositol-anchored protein in the plasma membrane.
AuthorsSheets ED, Lee GM, Simson R, Jacobson K
JournalBiochemistry
PubMed ID9376349
'Glycosylphosphatidylinositol (GPI)-anchored proteins participate in many cell surface functions; however, the molecular associations of these lipid-linked proteins within the plasma membrane are not well understood. Recent biochemical analyses of detergent insoluble membrane fractions have suggested that GPI-anchored proteins may be associated with glycosphingolipid (GSL)-enriched domains that also contain cholesterol and ... More
Lipid rafts reconstituted in model membranes.
AuthorsDietrich C, Bagatolli LA, Volovyk ZN, Thompson NL, Levi M, Jacobson K, Gratton E
JournalBiophys J
PubMed ID11222302
'One key tenet of the raft hypothesis is that the formation of glycosphingolipid- and cholesterol-rich lipid domains can be driven solely by characteristic lipid-lipid interactions, suggesting that rafts ought to form in model membranes composed of appropriate lipids. In fact, domains with raft-like properties were found to coexist with fluid ... More
Single-particle tracking of murine polyoma virus-like particles on live cells and artificial membranes.
AuthorsEwers H, Smith AE, Sbalzarini IF, Lilie H, Koumoutsakos P, Helenius A
JournalProc Natl Acad Sci U S A
PubMed ID16219700
'The lateral mobility of individual murine polyoma virus-like particles (VLPs) bound to live cells and artificial lipid bilayers was studied by single fluorescent particle tracking using total internal reflection fluorescence microscopy. The particle trajectories were analyzed in terms of diffusion rates and modes of motion as described by the moment ... More
Condensed complexes, rafts, and the chemical activity of cholesterol in membranes.
AuthorsRadhakrishnan A, Anderson TG, McConnell HM
JournalProc Natl Acad Sci U S A
PubMed ID11050164
'Epifluorescence microscopy studies of mixtures of phospholipids and cholesterol at the air-water interface often exhibit coexisting liquid phases. The properties of these liquids point to the formation of "condensed complexes" between cholesterol and certain phospholipids, such as sphingomyelin. It is found that monolayers that form complexes can incorporate a low ... More
The location of fluorescence probes with charged groups in model membranes.
AuthorsKachel K, Asuncion-Punzalan E, London E
JournalBiochim Biophys Acta
PubMed ID9814853
'The location of commonly used charged fluorescent membrane probes in membranes was determined in order to: (1) investigate the relationship between the structure of hydrophobic molecules and their depth within membranes; and (2) aid interpretation of experiments in which these fluorescent probes are used to examine membrane structure. Membrane depth ... More
Measuring the adsorption of Fatty acids to phospholipid vesicles by multiple fluorescence probes.
AuthorsSimard JR, Kamp F, Hamilton JA,
JournalBiophys J
PubMed ID18296488
'Fatty acids (FA) are important nutrients that the body uses to regulate the storage and use of energy resources. The predominant mechanism by which long-chain fatty acids enter cells is still debated widely as it is unclear whether long-chain fatty acids require protein transporters to catalyze their transmembrane movement. We ... More
Characterizing submicron vesicles with wavelength-resolved fluorescence in flow cytometry.
AuthorsFuller RR, Sweedler JV
JournalCytometry
PubMed ID8891444
'Individual synthetic vesicles 0.1-1.0 micron in diameter are sized by using single-particle fluorescence emission spectra obtained in a custom sheath flow cell with an imaging spectrograph and a charge-coupled device. Data are acquired at 1 Hz, with limits of detection (3 sigma) less than 6.0 x 10(3) and 1.0 x ... More
Novel Fusogenic Liposomes for Fluorescent Cell Labeling and Membrane Modification.
AuthorsCsisza´r A, Hersch N, Dieluweit S, Biehl R, Merkel R, Hoffmann B,
JournalBioconjug Chem
PubMed ID20184308
'Efficient delivery of biomolecules into membranes of living cells as well as cell surface modifications are major biotechnological challenges. Here, novel liposome systems based on neutral and cationic lipids in combination with lipids modified by aromatic groups are introduced for such applications. The fusion efficiency of these liposome systems was ... More
Morphology of proteoliposomes containing fluorescein-phosphatidylethanolamine reconstituted with native and subunit III-depleted cytochrome c oxidase.
AuthorsMüller M, Azzi A
JournalJ Bioenerg Biomembr
PubMed ID3007450
'Beef heart cytochrome c oxidase was reconstituted in asolectin liposomes containing the pH indicator fluorescein-phosphatidylethanolamine (FPE) by the cholate-dialysis procedure. The influence of FPE on the asolectin liposome size and of the removal of subunit III from the complex on its incorporation into liposomes was analyzed by freeze-fracture electron microscopy. ... More
The movement of fluorescent endocytic tracers in Plasmodium falciparum infected erythrocytes.
AuthorsHaldar K, Uyetake L
JournalMol Biochem Parasitol
PubMed ID1371847
'The fluorescent lipophilic probe 1,1''-dihexadecyl-3-3''-3-3''- tetramethylindocarbocyanine (diIC16) inserted in the red cell surface, functioned as a non-exchangeable lipid marker which was not metabolised or toxic in plasmodial cultures. Invasion by Plasmodium falciparum resulted in the internalisation of the lipid, suggesting the uptake of red cell membrane components during parasite entry. ... More
Deoxygenation affects fluorescence photobleaching recovery measurements of red cell membrane protein lateral mobility.
AuthorsCorbett JD, Cho MR, Golan DE
JournalBiophys J
PubMed ID8130343
'We have used the fluorescence photobleaching recovery technique to study the dependence on oxygen tension of the lateral mobility of fluorescently labeled band 3, the phospholipid analogue fluorescein phosphatidylethanolamine, and glycophorins in normal red blood cell membranes. Band 3 protein and sialic acid moieties on glycophorins were labeled specifically with ... More
Monopalmitoylphosphatidylcholine incorporation into human erythrocyte ghost membranes causes protein and lipid immobilization and cholesterol depletion.
AuthorsGolan DE, Furlong ST, Brown CS, Caulfield JP
JournalBiochemistry
PubMed ID3401442
'The effects of lysophosphatidylcholine (lysoPC) on human erythrocyte (RBC) ghost morphology, transmembrane protein and lipid lateral mobilities, and membrane lipid composition were studied in order to elucidate mechanisms by which lysoPC immobilizes ghost membrane components [Golan, D. E., Brown, C. S., Cianci, C. M. L., Furlong, S. T., & Caulfield, ... More
Intracellular translocation and metabolism of a fluorescent phosphatidic acid analogue in cultured fibroblasts.
AuthorsPagano RE, Longmuir KJ, Martin OC
JournalJ Biol Chem
PubMed ID6822547
'We have investigated the metabolism and intracellular translocation of a fluorescent derivative of phosphatidic acid, 1-acyl-2-[(N-4-nitrobenzo-2-oxa-1,3-diazole)aminocaproyl] phosphatidic acid (C6-NBD-PA), and its metabolites, in Chinese hamster fibroblasts. This derivative is rapidly transferred from phospholipid vesicles to cells at 2 degrees C, and results in fluorescent labeling of the mitochondria, endoplasmic reticulum, ... More
Interaction of free fatty acids with phospholipid bilayers.
AuthorsLangner M, Isac T, Hui SW
JournalBiochim Biophys Acta
PubMed ID7794957
'The partition of free fatty acids (FFA) to egg-phosphatidylcholine (egg-PC) and egg-phosphatidylethanolamine (egg-PE) vesicles was studied. Upon the addition of FFA to the suspension of vesicles, the pH of the aqueous phase changed depending on the length and saturation of the FFA hydrocarbon chain, as well as on the vesicle ... More
The effect of the lipid bilayer state on fluorescence intensity of fluorescein-PE in a saturated lipid bilayer.
AuthorsLangner M, Pruchnik H, Kubica K
JournalZ Naturforsch [C]
PubMed ID10928554
'Fluorescein-PE is a fluorescence probe that is used as a membrane label or a sensor of surface associated processes. Fluorescein-PE fluorescence intensity depends not only on bulk pH, but also on the local electrostatic potential, which affects the local membrane interface proton concentration. The pH sensitivity and hydrophilic character of ... More
The integral membrane protein, ponticulin, acts as a monomer in nucleating actin assembly.
AuthorsChia CP, Shariff A, Savage SA, Luna EJ
JournalJ Cell Biol
PubMed ID8432731
'Ponticulin, an F-actin binding transmembrane glycoprotein in Dictyostelium plasma membranes, was isolated by detergent extraction from cytoskeletons and purified to homogeneity. Ponticulin is an abundant membrane protein, averaging approximately 10(6) copies/cell, with an estimated surface density of approximately 300 per microns2. Ponticulin solubilized in octylglucoside exhibited hydrodynamic properties consistent with ... More
Regulation mechanism of the lateral diffusion of band 3 in erythrocyte membranes by the membrane skeleton.
AuthorsTomishige M, Sako Y, Kusumi A
JournalJ Cell Biol
PubMed ID9722611
'Mechanisms that regulate the movement of a membrane spanning protein band 3 in erythrocyte ghosts were investigated at the level of a single or small groups of molecules using single particle tracking with an enhanced time resolution (0.22 ms). Two-thirds of band 3 undergo macroscopic diffusion: a band 3 molecule ... More
Lateral proton conduction at a lipid/water interface. Its modulation by physical parameters. Experimental and mathematical approaches.
AuthorsPrats M, Tocanne JF, Teissié J
JournalEur J Biochem
PubMed ID2988956
'Fast lateral proton conduction along the lipid/water interface has recently been experimentally demonstrated in our laboratory [Teissié, J., Prats, M., Soucaille, P. & Tocanne, J.F. (1985) Proc. Natl Acad. Sci. USA, in the press]. The present study gives a more precise description of the way various physical parameters can affect ... More
Spatial organization of bacteriorhodopsin in model membranes. Light-induced mobility changes.
AuthorsKahya N, Wiersma DA, Poolman B, Hoekstra D
JournalJ Biol Chem
PubMed ID12167614
'Bacteriorhodopsin is a proton-transporting membrane protein in Halophilic archaea, and it is considered a prototype of membrane transporters and a model for G-protein-coupled receptors. Oligomerization of the protein has been reported, but it is unknown whether this feature is correlated with, for instance, light activation. Here, we have addressed this ... More
Control of band 3 lateral and rotational mobility by band 4.2 in intact erythrocytes: release of band 3 oligomers from low-affinity binding sites.
AuthorsGolan DE, Corbett JD, Korsgren C, Thatte HS, Hayette S, Yawata Y, Cohen CM
JournalBiophys J
PubMed ID8785311
'Band 4.2 is a human erythrocyte membrane protein of incompletely characterized structure and function. Erythrocytes deficient in band 4.2 protein were used to examine the functional role of band 4.2 in intact erythrocyte membranes. Both the lateral and the rotational mobilities of band 3 were increased in band 4.2-deficient erythrocytes ... More
UV immobilized phospholipid bilayers.
AuthorsUzgiris EE
JournalBiochem Biophys Res Commun
PubMed ID3619917
'Dinitrophenyl phosphotidylethanolamine-containing bilayers have been immobilized on carbon-shadowed support films by UV irradiation of the first monolayer transferred to the support film. The immobilized bilayer is capable of allowing bound protein (anti-DNP antibody) to organize into 2-D arrays in the presence of organic solvents such as acetonitrile and dilute concentrations ... More
Intramolecular distances within the Ca(2+)-ATPase from sarcoplasmic reticulum as estimated through fluorescence energy transfer between probes.
'Fluorescence energy transfer measurements have been carried out to estimate intramolecular distances between probes bound to Ca(2+)-transporting ATPase (Ca(2+)-ATPase) as well as distances between these probes and the phospholipid headgroup. The nucleotide binding site was monitored by using 1,N6-ethenoadenosine 5''-triphosphate, a fluorescent analogue of ATP, and also by labelling Lys515 ... More
Localization of the hinge region of the Ca(2+)-ATPase of sarcoplasmic reticulum using resonance energy transfer.
AuthorsBaker KJ, East JM, Lee AG
JournalBiochim Biophys Acta
PubMed ID8204650
'The Ca(2+)-ATPase of skeletal muscle sarcoplasmic reticulum can be labelled at Cys-670 and Cys-674 with 5-[[2-[(iodoacetyl) amino]ethyl]amino]naphthalene-1-sulphonic acid (IAEDANS). Resonance energy transfer has been used to measure the distance between Cys-670/Cys-674 and Glu-439 labelled with 5-(bromomethyl)fluorescein as 40 A. The height of Cys-670/Cys-674 above the phospholipid/water interface has been measured ... More
Green tea polyphenol epigallocatechin-3-gallate protects HepG2 cells against CYP2E1-dependent toxicity.
AuthorsJimenez-Lopez JM, Cederbaum AI
JournalFree Radic Biol Med
PubMed ID15036355
'Chronic ethanol consumption causes oxidative damage in the liver, and induction of cytochrome P450 2E1 (CYP2E1) is one pathway involved in oxidative stress produced by ethanol. The hepatic accumulation of iron and polyunsaturated fatty acids significantly contributes to ethanol hepatotoxicity in the intragastric infusion model of ethanol treatment. The objective ... More
Use of a fluorescein derivative of phosphatidylethanolamine as a pH probe at water/lipid interfaces.
AuthorsSoucaille P, Prats M, Tocanne JF, Teissié J
JournalBiochim Biophys Acta
PubMed ID3128325
'A fluorescent indicator for the determination of pH in the vicinity of water/lipid interfaces was produced by the covalent linkage of fluoresceinisothiocyanate to Escherichia coli phosphatidylethanolamine. When embedded in monolayers spread on an air/water interface, its apparent pK was shown to be only slightly affected by the nature of the ... More
The use of fluorescein-dipalmitoylphosphatidylethanolamine for measuring pH-changes in the internal compartment of phospholipid vesicles.
AuthorsThelen M, Petrone G, O'Shea PS, Azzi A
JournalBiochim Biophys Acta
PubMed ID6743649
'The synthesis and characterisation of fluorescein-phosphatidylethanolamine (FPE) is described. The effects of dielectric constant, ionic strength and ambient pH upon the optical absorbance properties of FPE are presented. It is shown that under appropriate conditions, FPE rapidly and quantitatively reports the pH of the aqueous bulk phases when incorporated into ... More
Resonance energy transfer assay of protein-mediated lipid transfer between vesicles.
AuthorsNichols JW, Pagano RE
JournalJ Biol Chem
PubMed ID6853520
Insertion of fluorescent phospholipids into the plasma membrane of a mammalian cell.
AuthorsStruck DK, Pagano RE
JournalJ Biol Chem
PubMed ID7372642
Lateral diffusion of lipids in model and natural membranes.
AuthorsTocanne JF, Dupou-Cézanne L, Lopez A
JournalProg Lipid Res
PubMed ID8022844
Lateral diffusion in planar lipid bilayers.
AuthorsFahey PF, Koppel DE, Barak LS, Wolf DE, Elson EL, Webb WW
JournalScience
PubMed ID831279
Direct measurements by fluorescence correlation spectroscopy of lateral diffusion coefficients of fluorescent lipid analogs in lipid bilaryer membranes indicate self-diffusion coefficients D greater than 10(-7) square centimeters per second for various lipid systems above their reported transition temperatures. Cholesterol in egg lecithin at mole ratio of 1 : 2 reduces ... More
Lipid lateral diffusion and membrane organization.
AuthorsTocanne JF, Dupou-Cézanne L, Lopez A, Tournier JF
JournalFEBS Lett
PubMed ID2680602
It is shown that investigating the lateral motion of lipids in biological membranes can provide useful information on membrane lateral organization. After labeling membranes with extrinsic or intrinsic lipophilic fluorescent probes, fluorescence recovery after photobleaching experiments strongly suggests that specialized cells like spermatozoa, eggs and epithelia exhibit surface membrane regionalization ... More
Flow cytometric determination of lipid peroxidation using fluoresceinated phosphoethanolamine.
AuthorsMaulik G, Salgia R, Makrigiorgos GM
JournalMethods Enzymol
PubMed ID12125379
In conclusion, we describe the use of fluor-DHPE as a flow cytometric probe to assess lipid peroxidation in erythrocytes. The stability and nonexchangeability of this probe make it suitable for monitoring lipid peroxidation in a particular cell type via flow cytometry, which can be done in the presence of other ... More
Positions of the sites labeled by N-cyclohexyl-N'-(4-dimethylamino-1-naphthyl)carbodiimide on the (Ca2+ + Mg2+)-ATPase.
AuthorsMunkonge F, East JM, Lee AG
JournalBiochim Biophys Acta
PubMed ID2521797
N-Cyclohexyl-N'-(4-dimethylamino-1-naphthyl)carbodiimide (NCD-4) labels (Ca2+ + Mg2+)-ATPase at Ca2+-protectable sites, believed to be at or near the two Ca2+ binding sites on the ATPase, and at nonspecific sites. The labeled ATPase has been reconstituted into lipid bilayers containing phosphatidylethanolamine labeled with fluorescein isothiocyanate. The distance between NCD-4 and fluorescein groups was ... More
Lateral proton conduction at a lipid/water interface. Effect of lipid nature and ionic content of the aqueous phase.
AuthorsPrats M, Tocanne JF, Teissie J
JournalEur J Biochem
PubMed ID3026808
Fast lateral proton conduction was observed along the lipid/water interface using a fluorescence technique. This conduction can be detected for a large number of lipids, both phospholipids and glycolipids. The efficiency of the proton transfer is dependent on the molecular packing of the host lipid at a given surface pressure. ... More
Surface-coupled proton exchange of a membrane-bound proton acceptor.
Proton-transfer reactions across and at the surface of biological membranes are central for maintaining the transmembrane proton electrochemical gradients involved in cellular energy conversion. In this study, fluorescence correlation spectroscopy was used to measure the local protonation and deprotonation rates of single pH-sensitive fluorophores conjugated to liposome membranes, and the ... More
How curved membranes recruit amphipathic helices and protein anchoring motifs.
Lipids and several specialized proteins are thought to be able to sense the curvature of membranes (MC). Here we used quantitative fluorescence microscopy to measure curvature-selective binding of amphipathic motifs on single liposomes 50-700 nm in diameter. Our results revealed that sensing is predominantly mediated by a higher density of ... More
Lipid vesicle adsorption versus formation of planar bilayers on solid surfaces.
AuthorsNollert P, Kiefer H, Jähnig F
JournalBiophys J
PubMed ID8534815
The absorption and spreading behavior of lipid vesicles composed of either palmitoyloleoylphosphatidylcholine (POPC) or Escherichia coli lipid upon contact with a glass surface was examined by fluorescence measurements. Fluorescently labeled lipids were used to determine 1) the amount of lipid adsorbed at the surface, 2) the extent of fusion of ... More
The position of the ATP binding site on the (Ca2+ + Mg2+)-ATPase.
AuthorsGutierrez-Merino C, Munkonge F, Mata AM, East JM, Levinson BL, Napier RM, Lee AG
JournalBiochim Biophys Acta
PubMed ID2949777
We present a convenient method to calculate the efficiency of fluorescence energy transfer in two-dimensional membrane systems. We apply it to the analysis of energy transfer between phospholipid molecules labelled with fluorescein and rhodamine groups, and of energy transfer in reconstituted membranes containing (Ca2+ + Mg2+)-ATPase purified from sarcoplasmic reticulum, ... More
Analysis of the structure and composition of individual lipoplex particles by flow fluorometry.
AuthorsPozharski EV, Macdonald RC
JournalAnal Biochem
PubMed ID15907868
A flow fluorometric approach to study cationic lipoid-DNA complexes is presented. The approach uses standard flow cytometry equipment and common fluorescent dyes (BODIPY and ethidium homodimer-2) to detect both lipoid and DNA content in individual particles. In addition, a procedure that allows determination of whether or not liposomes remain intact ... More
For H(+) transport between protein pumps, lateral diffusion along membrane surfaces represents the most efficient pathway. Along lipid bilayers, we measured a diffusion coefficient of 5.8 x 10(-5) cm(2) s(-1). It is too large to be accounted for by vehicle diffusion, considering proton transport by acid carriers. Such a speed ... More
Synthesis, characterization, and application of fluorescence sensing lipobeads for intracellular pH measurements.
AuthorsMcNamara KP, Nguyen T, Dumitrascu G, Ji J, Rosenzweig N, Rosenzweig Z
JournalAnal Chem
PubMed ID11476221
This paper describes the synthesis and characterization of micrometric phospholipid-coated polystyrene particles, named lipobeads, with pH-sensing capability and their application for intracellular pH measurements in murine macrophages. The phospholipids used to coat the particles are labeled with fluorescein (a pH-sensitive dye) and tetramethylrhodamine (a pH-insensitive dye), which serves as a ... More
Detecting proton flux across chromatophores driven by F0F1-ATPase using N-(fluorescein-5-thiocarbamoyl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt.
AuthorsYuanbo C, Fan Z, Jiachang Y
JournalAnal Biochem
PubMed ID16043113
N-(Fluorescein-5-thiocarbamoyl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt (F-DHPE) is a lipid fluorescence dye sensitive to pH changes and is used in this study for detecting proton flux through F0F1-ATPase within chromatophores driven by ATP hydrolysis. F-DHPE is easily labeled to the outer surface of chromatophores. In the range of pH 7.0 to 9.0, fluorescence ... More
Signals from the spindle midzone are required for the stimulation of cytokinesis in cultured epithelial cells.
AuthorsCao LG, Wang YL
JournalMol Biol Cell
PubMed ID8688554
The interaction between the mitotic spindle and the cellular cortex is thought to play a critical role in stimulating cell cleavage. However, little is understood about the nature of such interactions, particularly in tissue culture cells. We have investigated the role of the spindle midzone in signaling cytokinesis by creating ... More
Proton translocation by a native and subunit III-depleted cytochrome c oxidase reconstituted into phospholipid vesicles. Use of fluorescein-phosphatidylethanolamine as an intravesicular pH indicator.
AuthorsThelen M, O'Shea PS, Petrone G, Azzi A
JournalJ Biol Chem
PubMed ID2982869
The existence of a proton pump associated with bovine cytochrome c oxidase (EC 1.9.3.1) has over the last few years been a matter of considerable dispute. In an attempt to resolve some of the problems with the measuring system we have synthesized fluorescein-phosphatidylethanolamine which when reconstituted with cytochrome c oxidase ... More
Evidence for conduction of protons along the interface between water and a polar lipid monolayer.
AuthorsTeissié J, Prats M, Soucaille P, Tocanne JF
JournalProc Natl Acad Sci U S A
PubMed ID2987914
Movements of H+ along the polar heads of phospholipids spread in monolayers were compared to movements of H+ in the aqueous subphase. The probe for detecting H+ movement along the monolayer was a pH-sensitive fluorescein chromophore covalently bound to the head group of phosphatidylethanolamine. The behavior of this probe was ... More
Measurements of interbilayer forces and protein adsorption on uncharged lipid bilayers displaying poly(ethylene glycol) chains.
AuthorsEfremova NV, Bondurant B, O'Brien DF, Leckband DE
JournalBiochemistry
PubMed ID10727239
Poly(ethylene glycol) (PEG)-stabilized liposomes were recently shown to exhibit differences in cell uptake that were linked to the liposome charge. To determine the differences and similarities between charged and uncharged PEG-decorated liposomes, we directly measured the forces between two supported, neutral bilayers with terminally grafted PEG chains. The measurements were ... More
Lipid diffusibility in the intact erythrocyte membrane.
AuthorsBloom JA, Webb WW
JournalBiophys J
PubMed ID6603237
The lateral diffusion of fluorescent lipid analogues in the plasma membrane of intact erythrocytes from man, mouse, rabbit, and frog has been measured by fluorescence photobleaching recovery (FPR). Intact cells from dystrophic, normoblastic, hemolytic, and spherocytotic mouse mutants; from hypercholesterolemic rabbits and humans; and from prenatal, neonatal, and juvenile mice ... More
The lateral diffusion of lipid probes in the surface membrane of Schistosoma mansoni.
AuthorsFoley M, MacGregor AN, Kusel JR, Garland PB, Downie T, Moore I
JournalJ Cell Biol
PubMed ID3745270
The technique of fluorescence recovery after photobleaching was used to measure the lateral diffusion of fluorescent lipid analogues in the surface membrane of Schistosoma mansoni. Our data reveal that although some lipids could diffuse freely others exhibited restricted lateral diffusion. Quenching of lipid fluorescence by a non-permeant quencher, trypan blue, ... More
High-generation polycationic dendrimers are unusually effective at disrupting anionic vesicles: membrane bending model.
AuthorsZhang ZY, Smith BD
JournalBioconjug Chem
PubMed ID11087328
The membrane disruption properties of high generation (G4 to G7) poly(amidoamine) (PAMAM) dendrimers are evaluated and compared to linear poly(lysine). The G6 and G7 dendrimers are unusually effective at inducing leaky fusion of anionic, large unilamellar vesicles, as determined by standard fluorescence assays for lipid mixing, leakage, and contents mixing. ... More
Cell spreading and lamellipodial extension rate is regulated by membrane tension.
AuthorsRaucher D, Sheetz MP
JournalJ Cell Biol
PubMed ID10629223
Cell spreading and motility require the extension of the plasma membrane in association with the assembly of actin. In vitro, extension must overcome resistance from tension within the plasma membrane. We report here that the addition of either amphiphilic compounds or fluorescent lipids that expanded the plasma membrane increased the ... More
Hydration-driven transport of deformable lipid vesicles through fine pores and the skin barrier.
AuthorsCevc G, Gebauer D
JournalBiophys J
PubMed ID12547782
We studied aggregate transport through semipermeable, nano-porous barriers experimentally and theoretically. By measuring and modeling the effect of hydration gradient across such barriers, spontaneous transbarrier transport of suitable lipid aggregates in vesicular form was proven to be driven by partial aggregate dehydration at the application site. By generalizing the Onsager ... More
Enhancement of molecular fluorescence near the surface of colloidal metal films.
AuthorsSokolov K, Chumanov G, Cotton TM
JournalAnal Chem
PubMed ID9751028
Fluorescence enhancement was studied on silver colloidal metal films (CMFs) using two systems: (1) Langmuir--Blodgett monolayers of fluorescein-labeled phospholipids separated from the surface of the films by spacer layers of octadecanoic acid and (2) biotin--fluorescein conjugates captured by avidin molecules adsorbed on top of a multilayer structure formed by alternating ... More
Comparison of reagents for shape analysis of fixed cells by automated fluorescence microscopy.
AuthorsElliott JT, Tona A, Plant AL
JournalCytometry A
PubMed ID12655652
BACKGROUND: Cell size and shape have been implicated as potentiators of intracellular signaling events and as indicators of abnormal cell behavior. Automated microscopy and image analysis can provide quantitative information about the size and shape of cultured cells, but it requires that the edge of a cell be clearly identified. ... More
Red cell membrane remodeling in sickle cell anemia. Sequestration of membrane lipids and proteins in Heinz bodies.
AuthorsLiu SC, Yi SJ, Mehta JR, Nichols PE, Ballas SK, Yacono PW, Golan DE, Palek J
JournalJ Clin Invest
PubMed ID8550846
In red cells from patients with sickle cell anemia, hemoglobin S denatures and forms Heinz bodies. Binding of Heinz bodies to the inner surface of the sickle cell membrane promotes clustering and colocalization of the membrane protein band 3, outer surface-bound autologous IgG and, to some extent, the membrane proteins ... More
Fluorescent liposome flow markers for microscale particle-image velocimetry.
AuthorsSingh AK, Cummings EB, Throckmorton DJ
JournalAnal Chem
PubMed ID11289418
Unilamellar liposomes carrying both encapsulated and surface-immobilized fluorophores have been synthesized as novel fluorescent markers to image flow profiles in microfabricated structures. The unilamellar liposomes were made with phospholipids and cholesterol by extrusion through a polycarbonate membrane. They contained carboxyfluorescein in the aqueous core and fluorescein-labeled lipids in the bilayer ... More
Lateral proton conduction in monolayers of phospholipids from extreme halophiles.
AuthorsTeissié J, Prats M, LeMassu A, Stewart LC, Kates M
JournalBiochemistry
PubMed ID2157482
Studies have been carried out on the lateral proton conductance properties of monolayers of the major and minor phospholipids of extremely halophilic archaebacteria, 2,3-diphytanyl-sn-glycero-1-phospho-3'-sn-glycerol 1'-phosphate (PGP) and 2,3-diphytanyl-sn-glycero-1-phospho-3'-sn-glycerol (PG), respectively, as well as on their respective deoxy analogues: 2,3-diphytanyl-sn-glycero-1-phospho-1'-propanediol 3'-phosphate (dPGP), 2,3-diphytanyl-sn-glycero-1-phospho-1'-1',3'-propanediol (dPG), and 2,3-diphytanyl-sn-glycero-1-phospho-1'-propanol (ddPG). Lateral proton conduction was ... More
Differing reactivities of human and guinea-pig complement on haptenized liposomes via the alternative pathway.
AuthorsOkada N, Yasuda T, Tsumita T, Okada H
JournalMol Immunol
PubMed ID6194429
Several haptens coupled to dipalmitoylphosphatidylethanolamine (DPPE) were inserted into the liposome membrane with a base composition of an equimolecular mixture of dimyristoylphosphatidylcholine (DMPC) and cholesterol (Chol). Haptens used were trinitrophenyl (TNP)-DPPE, TNP-aminocaproyl (TNP-Cap)-DPPE, dinitrophenyl (DNP)-DPPE, DNP-aminocaproyl (DNP-Cap)-DPPE, fluoresceinthiocarbamyl (Fl)-DPPE, azobenzenarsonate-tyrosyl (ABA-Tyr)-DPPE, dansyl (DNS)-DPPE, dabsyl (DABS)-DPPE, dithiopyridyl (DTP)-DPPE and maleimidobenzoyl (MB)-DPPE. ... More
Unconfined lateral diffusion and an estimate of pericellular matrix viscosity revealed by measuring the mobility of gold-tagged lipids.
AuthorsLee GM, Zhang F, Ishihara A, McNeil CL, Jacobson KA
JournalJ Cell Biol
PubMed ID8416991
Nanovid (video-enhanced) microscopy was used to determine whether lateral diffusion in the plasma membrane of colloidal gold-tagged lipid molecules is confined or is unrestricted. Confinement could be produced by domains within the plane of the plasma membrane or by filamentous barriers within the pericellular matrix. Fluorescein-phosphatidylethanolamine (F1-PE), incorporated into the ... More
Class I major histocompatibility complex proteins diffuse isotropically on immune interferon-activated endothelial cells despite anisotropic cell shape and cytoskeletal organization: application of fluorescence photobleaching recovery with an elliptical beam.
AuthorsStolpen AH, Pober JS, Brown CS, Golan DE
JournalProc Natl Acad Sci U S A
PubMed ID3126497
Interferon gamma induces striking phenotypic alterations in confluent cultures of human vascular endothelial cells (HEC), including cell shape change from polygonal to elongated and cytoskeletal actin rearrangement from dense peripheral bands to longitudinal bundles of stress fibers. Since many transmembrane proteins, including class I major histocompatibility complex (MHC) proteins, interact ... More
Tumor necrosis factor and immune interferon act in concert to slow the lateral diffusion of proteins and lipids in human endothelial cell membranes.
AuthorsStolpen AH, Golan DE, Pober JS
JournalJ Cell Biol
PubMed ID3138247
Vascular endothelial surface-related activities may depend on the lateral mobility of specific cell surface macromolecules. Previous studies have shown that cytokines induce changes in the morphology and surface antigen composition of vascular endothelial cells in vitro and at sites of immune and inflammatory reactions in vivo. The effects of cytokines ... More
Evidence for the formation of microdomains in liquid crystalline large unilamellar vesicles caused by hydrophobic mismatch of the constituent phospholipids.
AuthorsLehtonen JY, Holopainen JM, Kinnunen PK
JournalBiophys J
PubMed ID8785334
The excimer-to-monomer fluorescence emission intensity ratio (IE/IM) of the fluorescent probe 1-palmitoyl-2-[(pyren-1-yl)]decanoyl-sn-glycero-3-phosphocholine (PPDPC, 1 mol%) was measured at 30 degrees C as a function of the thickness of fluid liposomal membranes composed of phosphatidylcholines (PCs) with homologous monounsaturated acyl chains of varying lengths N (= number of carbon atoms). Upon ... More
Transverse location of the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene in model lipid bilayer membrane systems by resonance excitation energy transfer.
AuthorsDavenport L, Dale RE, Bisby RH, Cundall RB
JournalBiochemistry
PubMed ID3931673
A fluorescent phospholipid derivative, the fluoresceinthiocarbamyl adduct of a natural phosphatidylethanolamine, has been synthesized and incorporated into sonicated single-bilayer vesicles of egg lecithin and dipalmitoyllecithin. The surface location of this probe has been confirmed by using extrinsic fluorescence quenching studies together with steady-state emission anisotropy measurements. Electronic excitation energy transfer ... More
Outer membrane monolayer domains from two-dimensional surface scanning resistance measurements.
AuthorsSuzuki K, Sterba RE, Sheetz MP
JournalBiophys J
PubMed ID10866970
Cellular plasma membranes have domains that are defined, in most cases, by cytoskeletal elements. The outer half of the bilayer may also contain domains that organize glycosylphosphatidylinositol (GPI)-linked proteins. To define outer membrane barriers, we measured the resistive force on membrane bound beads as they were scanned across the plasma ... More
Quenching of fluorescein-conjugated lipids by antibodies. Quantitative recognition and binding of lipid-bound haptens in biomembrane models, formation of two-dimensional protein domains and molecular dynamics simulations.
AuthorsAhlers M, Grainger DW, Herron JN, Lim K, Ringsdorf H, Salesse C
JournalBiophys J
PubMed ID1420916
Three model biomembrane systems, monolayers, micelles, and vesicles, have been used to study the influence of chemical and physical variables of hapten presentation at membrane interfaces on antibody binding. Hapten recognition and binding were monitored for the anti-fluorescein monoclonal antibody 4-4-20 generated against the hapten, fluorescein, in these membrane models ... More
Low density lipoproteins bound to Schistosoma mansoni do not alter the rapid lateral diffusion or shedding of lipids in the outer surface membrane.
AuthorsCaulfield JP, Chiang CP, Yacono PW, Smith LA, Golan DE
JournalJ Cell Sci
PubMed ID1757500
Schistosomula of Schistosoma mansoni bind human low density lipoproteins (LDL) in a concentration-dependent and saturable manner. The bound LDL could provide phospholipids and sterol to the worm, which cannot synthesize sterol de novo and lacks acyl chain-modifying capability. Here we have used three phospholipid analogues to explore the effect of ... More
Direct observation of brownian motion of lipids in a membrane.
AuthorsLee GM, Ishihara A, Jacobson KA
JournalProc Natl Acad Sci U S A
PubMed ID1712486
Nanovid microscopy, which uses 30- to 40-nm colloidal gold probes combined with video-enhanced contrast, can be used to examine random and directed movements of individual molecules in the plasma membrane of living cells. To validate the technique in a model system, the movements of lipid molecules were followed in a ... More
Binding of cross-linked glycosylphosphatidylinositol-anchored proteins to discrete actin-associated sites and cholesterol-dependent domains.
AuthorsSuzuki K, Sheetz MP
JournalBiophys J
PubMed ID11566789
The mechanism by which cross-linked glycosylphosphatidylinositol (GPI)-anchored proteins are immobilized has been a mystery because both the binding to a transmembrane protein and attachment to a rigid cytoskeleton are needed. Using laser tweezers surface scanning resistance (SSR) technology, we obtained physical evidence for cross-linked GPI-anchored protein, Qa-2, binding to a ... More
Fluorescent phospholipids preferentially accumulate in sub-tegumental cells of schistosomula of Schistosoma mansoni.
AuthorsFurlong ST, Thibault KS, Rogers RA
JournalJ Cell Sci
PubMed ID1478974
Schistosomes do not make sterols or fatty acids de novo and thus require host lipids for survival. The acquisition of host lipids may also be an important factor in the schistosome's defense from host immunity; however, little is known about the regulation of this process. Here we have examined binding ... More
Schistosomula of Schistosoma mansoni use lysophosphatidylcholine to lyse adherent human red blood cells and immobilize red cell membrane components.
AuthorsGolan DE, Brown CS, Cianci CM, Furlong ST, Caulfield JP
JournalJ Cell Biol
PubMed ID3745271
Human red blood cells (RBCs) adhere to and are lysed by schistosomula of Schistosoma mansoni. We have investigated the mechanism of RBC lysis by comparing the dynamic properties of transmembrane protein and lipid probes in adherent ghost membranes with those in control RBCs and in RBCs treated with various membrane ... More
Nitric oxide enhances the manganese superoxide dismutase-dependent suppression of proliferation in HT-1080 fibrosarcoma cells.
AuthorsMelendez JA, Melathe RP, Rodriguez AM, Mazurkiewicz JE, Davies KJ
JournalCell Growth Differ
PubMed ID10511315
The overexpression of manganese superoxide dismutase (MnSOD), an enzyme that catalyzes the removal of superoxide (O2*-) from the mitochondria, has been shown to be closely associated with tumor regression in vivo and loss of the malignant phenotype in vitro. To investigate the mechanism by which MnSOD overexpression mediates this reversal, ... More
Tracking movements of lipids and Thy1 molecules in the plasmalemma of living fibroblasts by fluorescence video microscopy with nanometer scale precision.
AuthorsHicks BW, Angelides KJ
JournalJ Membr Biol
PubMed ID7658460
The lateral diffusion of 100 nm fluorescent latex microspheres (FS) bound to either N-biotinyl-phosphatidyl-ethanolamine or the glycosylphosphatidylinositol-linked protein Thy1 were monitored in the plasmalemma of primary rat fibroblasts by single particle tracking of FS centroids from digital fluorescence micrographs. A silicon intensified target camera was found to be superior to ... More
Interaction of a nonspecific wheat lipid transfer protein with phospholipid monolayers imaged by fluorescence microscopy and studied by infrared spectroscopy.
AuthorsSubirade M, Salesse C, Marion D, Pézolet M
JournalBiophys J
PubMed ID8519997
The interaction of a nonspecific wheat lipid transfer protein (LTP) with phospholipids has been studied using the monolayer technique as a simplified model of biological membranes. The molecular organization of the LTP-phospholipid monolayer has been determined by using polarized attenuated total internal reflectance infrared spectroscopy, and detailed information on the ... More
Reconstitution of membrane proteins into giant unilamellar vesicles via peptide-induced fusion.
AuthorsKahya N, Pécheur EI, de Boeij WP, Wiersma DA, Hoekstra D
JournalBiophys J
PubMed ID11509360
In this work, we present a protocol to reconstitute membrane proteins into giant unilamellar vesicles (GUV) via peptide-induced fusion. In principle, GUV provide a well-defined lipid matrix, resembling a close-to-native state for biophysical studies, including optical microspectroscopy, of transmembrane proteins at the molecular level. Furthermore, reconstitution in this manner would ... More
Characterization of the sequence of interactions of the fusion domain of the simian immunodeficiency virus with membranes. Role of the membrane dipole potential.
AuthorsCladera J, Martin I, Ruysschaert JM, O'Shea P
JournalJ Biol Chem
PubMed ID10514478
The simian immunodeficiency virus fusion peptide constitutes a 12-residue N-terminal segment of the gp32 protein that is involved in the fusion between the viral and cellular membranes, facilitating the penetration of the virus in the host cell. Simian immunodeficiency virus fusion peptide is a hydrophobic peptide that in Me(2)SO forms ... More
Semi-Automated Cell Panning for Efficient Isolation of FGFR3-Targeting Antibody.