Use of resonance energy transfer to study the kinetics of amphiphile transfer between vesicles.
AuthorsNichols JW, Pagano RE
JournalBiochemistry
PubMed ID7082641
Phospholipid translocation and miltefosine potency require both L. donovani miltefosine transporter and the new protein LdRos3 in Leishmania parasites.
AuthorsPérez-Victoria FJ, Sánchez-Cañete MP, Castanys S, Gamarro F
JournalJ Biol Chem
PubMed ID16785229
'The antitumor drug miltefosine has been recently approved as the first oral drug active against visceral leishmaniasis. We have previously identified the L. donovani miltefosine transporter (LdMT) as a P-type ATPase involved in phospholipid translocation at the plasma membrane of Leishmania parasites. Here we show that this protein is essential ... More
Membrane domains containing phosphatidylserine and substrate can be important for the activation of protein kinase C.
AuthorsYang L, Glaser M
JournalBiochemistry
PubMed ID7531492
'The relationship between lipid domains and enzyme activity was studied via the direct visualization and quantitation of domains by fluorescence digital imaging microscopy. The substrate used in these experiments was a basic peptide derived from a prominent cellular substrate (MARCKS) of protein kinase C. The MARCKS peptide and phosphatidylserine, which ... More
Localization of phospholipase D in detergent-insoluble, caveolin-rich membrane domains. Modulation by caveolin-1 expression and caveolin-182-101.
AuthorsCzarny M, Lavie Y, Fiucci G, Liscovitch M
JournalJ Biol Chem
PubMed ID9915802
'The activation of cellular phospholipase D (PLD) is implicated in vesicular trafficking and signal transduction. Two mammalian PLD forms, designated PLD1 and PLD2, have been cloned, but their cellular localization and function are not fully understood. Here, we report that in HaCaT human keratinocytes, as well as other cell lines, ... More
Formation of membrane domains created during the budding of vesicular stomatitis virus. A model for selective lipid and protein sorting in biological membranes.
AuthorsLuan P, Yang L, Glaser M
JournalBiochemistry
PubMed ID7543280
'Vesicular stomatitis virus buds from domains of the plasma membrane that have a unique protein and lipid composition. Fluorescence digital imaging microscopy and resonance energy transfer were used to determine how the two viral envelope-associated proteins, the G and the M proteins, could alter the lateral distribution of lipids in ... More
Fluorescence microscopy of phospholipid monolayer phase transitions.
AuthorsWeis RM
JournalChem Phys Lipids
PubMed ID2054906
'Over many years, a detailed picture of the phase transitions in phospholipid monolayers at the air-water interface has been constructed from extensive studies of the force-area, viscoelastic and surface potential properties of phospholipid monolayers, yet the microscopic nature of the transitions has remained obscure. Recent investigations have focused specifically on ... More
Lateral organization and domain formation in a two-component lipid membrane system.
'The thermodynamic phase behavior and lateral lipid membrane organization of unilamellar vesicles made from mixtures of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and 1,2 distearoyl-sn-glycero-3-phosphocholine (DSPC) were investigated by fluorescence resonance energy transfer (FRET) as a function of temperature and composition. This was done by incorporating a headgroup-labeled lipid donor (NBD-DPPE) and acceptor (N-Rh-DPPE) ... More
Transmembrane movement of diether phospholipids in human erythrocytes and human fibroblasts.
'We have synthesized spin-labeled (SL) and fluorescently labeled diacyl, 1-alkyl-2-acyl-, and di-alkyl glycerophospholipids. The sn-2 chain was a short chain with either a nitroxide group or a 7-nitro-2, 1,3-benzoxadiazol-4-yl (NBD). After incorporation in the exoplasmic leaflet of human erythrocytes, we found that SL-phosphatidylcholine (PC) redistributed very slowly across the plasma ... More
Facilitated phosphatidylcholine flip-flop across erythrocyte membranes using low molecular weight synthetic translocases.
AuthorsBoon JM, Smith BD
JournalJ Am Chem Soc
PubMed ID11427044
'The transmembrane distribution of phospholipids plays an important regulatory role in human erythrocytes. Membrane-bound translocase enzymes maintain an asymmetric phospholipid distribution across the membrane monolayers by promoting transmembrane diffusion or flip-flop. Mechanistic understanding of the flip-flop process is weak at the molecular level. Recently, we discovered that amide and sulfonamide ... More
Lateral diffusion measurement at high spatial resolution by scanning microphotolysis in a confocal microscope.
AuthorsKubitscheck U, Wedekind P, Peters R
JournalBiophys J
PubMed ID7811951
'Fluorescence photobleaching methods have been widely used to study diffusion processes in the plasma membrane of single living cells and other membrane systems. Here we describe the application of a new photobleaching technique, scanning microphotolysis. Employing a recently developed extension module to a commercial confocal microscope, an intensive laser beam ... More
Up-regulation of phospholipase D activity induced by overexpression of protein kinase C-alpha. Studies in intact Swiss/3T3 cells and in detergent-solubilized membranes in vitro.
AuthorsEldar H, Ben-Av P, Schmidt US, Livneh E, Liscovitch M
JournalJ Biol Chem
PubMed ID8509396
'The role of protein kinase C in the mechanism of phospholipase D activation by platelet-derived growth factor and 12-O-tetradecanoylphorbol-13-acetate was studied in Swiss/3T3 fibroblasts that overexpress protein kinase C-alpha. Production of [3H]phosphatidylpropanol (specific product of the phospholipase D-catalyzed transphosphatidylation reaction) was determined in cells which were prelabeled with [3H]oleic acid. ... More
gamma-cyclodextrins greatly enhance translocation of hydrophobic fluorescent phospholipids from vesicles to cells in culture. Importance of molecular hydrophobicity in phospholipid trafficking studies.
AuthorsTanhuanpää K, Somerharju P
JournalJ Biol Chem
PubMed ID10585403
'Short-chain, fluorescent derivatives are commonly used to investigate intracellular phospholipid trafficking. However, their use can yield misleading results because they, unlike the native species, can rapidly distribute between organelles due to their low hydrophobicity. On the other hand, hydrophobic derivatives are very difficult to introduce to cells and thus have ... More
Identification of a novel family of nonclassic yeast phosphatidylinositol transfer proteins whose function modulates phospholipase D activity and Sec14p-independent cell growth.
AuthorsLi X, Routt SM, Xie Z, Cui X, Fang M, Kearns MA, Bard M, Kirsch DR, Bankaitis VA
JournalMol Biol Cell
PubMed ID10848624
'Yeast phosphatidylinositol transfer protein (Sec14p) is essential for Golgi function and cell viability. We now report a characterization of five yeast SFH (Sec Fourteen Homologue) proteins that share 24-65% primary sequence identity with Sec14p. We show that Sfh1p, which shares 64% primary sequence identity with Sec14p, is nonfunctional as a ... More
Cell-cell adhesion mediated by a receptor-like protein tyrosine phosphatase.
AuthorsGebbink MF, Zondag GC, Wubbolts RW, Beijersbergen RL, van Etten I, Moolenaar WH
JournalJ Biol Chem
PubMed ID8393854
'Receptor-like protein tyrosine phosphatases (receptor-PTPs) represent a novel family of transmembrane proteins that are thought to play important roles in cellular regulation. They consist of a cytoplasmic catalytic region, a single transmembrane segment and an extracellular, putative ligand-binding domain, but the nature of their physiological ligands is unknown. We have ... More
Calcium/phosphate-induced immobilization of fluorescent phosphatidylserine in synthetic bilayer membranes: inhibition of lipid transfer between vesicles.
AuthorsTanaka Y, Schroit AJ
JournalBiochemistry
PubMed ID3707938
'Resonance energy transfer between 4-nitro-2,1,3-benzoxadiazole (NBD) acyl chain labeled phospholipid analogues and (lissamine) rhodamine B labeled phosphatidylethanolamine was used to monitor the rate of NBD-labeled lipid transfer between a variety of small unilamellar donor vesicles and dioleoylphosphatidylcholine (DOPC) acceptor vesicles. In the presence of appropriate concentrations of Ca2+ and phosphate, ... More
Transverse redistribution of phospholipids during human platelet activation: evidence for a vectorial outflux specific to aminophospholipids.
AuthorsGaffet P, Bettache N, Bienvenüe A
JournalBiochemistry
PubMed ID7756307
'The redistribution kinetics of phospholipids during human platelet activation by calcium ionophore were investigated to determine the specificity of the observed phospholipid outflux [Bassé et al. (1993) Biochemistry 32, 2337]. (1) Two double-labeling experiments were performed with a combination of equal amounts of spin- and fluorescently-labeled phosphatidylserine and phosphatidylcholine. During ... More
Lipid composition determines the effects of arbutin on the stability of membranes.
AuthorsHincha DK, Oliver AE, Crowe JH
JournalBiophys J
PubMed ID10512822
'Arbutin (hydroquinone-beta-D-glucopyranoside) is an abundant solute in the leaves of many freezing- or desiccation-tolerant plants. Its physiological role in plants, however, is not known. Here we show that arbutin protects isolated spinach (Spinacia oleracea L.) thylakoid membranes from freeze-thaw damage. During freezing of liposomes, the presence of only 20 mM ... More
Inhibition of in vitro endosomal vesicle fusion activity by aminoglycoside antibiotics.
AuthorsJones AT, Wessling-Resnick M
JournalJ Biol Chem
PubMed ID9737996
'The effects of two aminoglycoside antibiotics, neomycin and Geneticin, on the endocytic pathway were studied using a cell-free assay that reconstitutes endosome-endosome fusion. Both drugs inhibit the rate and extent of endosome fusion in a dose-dependent manner with IC50 values of approximately 45 microM and approximately 1 mM, respectively. Because ... 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
Pseudoapoptosis induced by brief activation of ATP-gated P2X7 receptors.
AuthorsMackenzie AB, Young MT, Adinolfi E, Surprenant A
JournalJ Biol Chem
PubMed ID15994333
'P2X7 receptors are ATP-gated ion channels primarily expressed on antigen-presenting immune cells where they play a role in the acute inflammatory response. These ion channels couple not only to influx of cations, including calcium, but also to rapid alterations in cell morphology (membrane blebbing, phosphatidylserine exposure, microvesicle shedding). These features ... More
Phosphatidylserine decarboxylase from Saccharomyces cerevisiae. Isolation of mutants, cloning of the gene, and creation of a null allele.
AuthorsTrotter PJ, Pedretti J, Voelker DR
JournalJ Biol Chem
PubMed ID8407984
'Phosphatidylserine decarboxylase plays a pivotal role in the synthesis of phospholipid by the mitochondria. The substrate phosphatidylserine is synthesized extramitochondrially and must be translocated to the mitochondria prior to decarboxylation. To understand the properties of the decarboxylase and exploit its unique topology to address basic questions of interorganelle cooperation in ... More
Use of spin-labeled and fluorescent lipids to study the activity of the phospholipid transfer protein from maize seedlings.
AuthorsGeldwerth D, de Kermel A, Zachowski A, Guerbette F, Kader JC, Henry JP, Devaux PF
JournalBiochim Biophys Acta
PubMed ID1851443
'The transfer of spin-labeled and fluorescent lipids between sonicated vesicles and different host membranes has been measured in the presence or absence of a phospholipid transfer protein purified from maize seedlings. It was found that the protein has little specificity towards the phospholipid head group and allows the transfer of ... More
Energy-dependent flip of fluorescence-labeled phospholipids is regulated by nutrient starvation and transcription factors, PDR1 and PDR3.
AuthorsHanson PK, Nichols JW
JournalJ Biol Chem
PubMed ID11136727
'The yeast Saccharomyces cerevisiae readily accumulates short-chain, fluorescent 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD)-labeled phosphatidylcholine and phosphatidylethanolamine at the nuclear envelope/endoplasmic reticulum and mitochondria. The net intracellular accumulation reflects the sum of their inwardly and outwardly directed transbilayer translocation across the plasma membrane (flip and flop, respectively). The rate of flop is negligible in ... More
Evidence for a Trypanosoma brucei lipoprotein scavenger receptor.
AuthorsGreen HP, Del Pilar Molina Portela M, St Jean EN, Lugli EB, Raper J
JournalJ Biol Chem
PubMed ID12401813
'African trypanosomes are lipid auxotrophs that live in the bloodstream of their human and animal hosts. Trypanosomes require lipoproteins in addition to other serum components in order to multiply under axenic culture conditions. Delipidation of the lipoproteins abrogates their capacity to support trypanosome growth. Both major classes of serum lipoproteins, ... More
Pyrularia thionin binding to and the role of tryptophan-8 in the enhancement of phosphatidylserine domains in erythrocyte membranes.
AuthorsWang F, Naisbitt GH, Vernon LP, Glaser M
JournalBiochemistry
PubMed ID8241114
'Pyrularia thionin is a small, strongly basic peptide which interacts readily with cellular and synthetic membranes. With cells it induces hemolysis, depolarizes the cellular membrane with an accompanying influx of Ca2+, and activates an endogenous phospholipase A2. Evidence points toward a binding site involving phosphatidylserine (PS). This study shows that ... More
Formation of asymmetric phospholipid membranes via spontaneous transfer of fluorescent lipid analogues between vesicle populations.
AuthorsPagano RE, Martin OC, Schroit AJ, Struck DK
JournalBiochemistry
PubMed ID7295659
'A method is presented for generating artificial lipid vesicles bearing an asymmetric distribution of either of the fluorescent lipid analogues 1-acyl-2-[6-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]caproyl]phosphatidylcholine or 1-acyl-2-[12[(7-nitro-2,1,3-benzoxadiazol-4-yl]amino]dodecanoyl]-phosphatidylcholine, in which the fluorescent lipid is located predominantly in either the outer or inner leaflet of the vesicle bilayer. The procedure is based on the observation that ... More
Binding of fluorescent phosphatidylcholine in amniotic fluid.
AuthorsAshwood ER, Chamberlain BV
JournalObstet Gynecol
PubMed ID3347421
'Fetal lung maturity can be predicted by the technique of fluorescence polarization using fluorescent phosphatidylcholine instead of diphenylhexatriene. To further elucidate the mechanism of this assay, we used high-speed centrifugation to isolate amniotic fluid lamellar bodies. Each of 75 amniotic fluid samples was separated into two fractions: a lamellar body ... More
Association of cytoplasmic free Ca2+ gradients with subcellular organelles.
AuthorsWahl M, Sleight RG, Gruenstein E
JournalJ Cell Physiol
PubMed ID1537888
'Previous investigations have identified gradients of intracellular free (Ca2+)i (Ca2+i) in the cytoplasm of human fibroblasts. In this study we have compared the spatial distribution of these gradients with the subcellular distribution of cytoplasmic organelles. Using the Ca(2+)-sensitive dye fura-2 and organelle-specific fluorescent dyes, we have found that the highest ... More
Phospholipid flippase activity of the reconstituted P-glycoprotein multidrug transporter.
AuthorsRomsicki Y, Sharom FJ
JournalBiochemistry
PubMed ID11389609
'The P-glycoprotein multidrug transporter acts as an ATP-powered efflux pump for a large variety of hydrophobic drugs, natural products, and peptides. The protein is proposed to interact with its substrates within the hydrophobic interior of the membrane. There is indirect evidence to suggest that P-glycoprotein can also transport, or "flip", ... More
Preferential distribution of the fluorescent phospholipid probes NBD-phosphatidylcholine and rhodamine-phosphatidylethanolamine in the exofacial leaflet of acetylcholine receptor-rich membranes from Torpedo marmorata.
AuthorsGutiérrez-Merino C, Bonini de Romanelli IC, Pietrasanta LI, Barrantes FJ
JournalBiochemistry
PubMed ID7718591
'The distribution of the two fluorescent phospholipid analogs across acetylcholine receptor (AChR)-rich membranes from Torpedo marmorata has been studied by a combination of nonradiative fluorescence resonance energy transfer using fluorescent lipid probes and quenching of their fluorescence with Co2+ and 2,4,6-trinitrobenzenesulfonic acid. The fluorescent lipid analogs were supplied to the ... More
Visualization of domain formation in the inner and outer leaflets of a phospholipid bilayer.
AuthorsHaverstick DM, Glaser M
JournalJ Cell Biol
PubMed ID3384848
'Large vesicles (5-10-micron in diameter) were formed in the presence of phospholipids fluorescently labeled on the acyl chain and visualized using a fluorescence microscope, charge-coupled-device camera and digital image processor. When such vesicles contained a fluorescent phosphatidic acid (PA) and were exposed to 2 mM CaCl2 or 0.5 mM PrCl3, ... More
A role for phospholipase D (Pld1p) in growth, secretion, and regulation of membrane lipid synthesis in yeast.
AuthorsSreenivas A, Patton-Vogt JL, Bruno V, Griac P, Henry SA
JournalJ Biol Chem
PubMed ID9642212
'The SEC14 gene encodes a phosphatidylinositol/phosphatidylcholine transfer protein essential for secretion and growth in yeast (1). Mutations (cki1, cct1, and cpt1) in the CDP-choline pathway for phosphatidylcholine synthesis suppress the sec14 growth defect (2), permitting sec14(ts) cki1, sec14(ts) cct1, and sec14(ts) cpt1 strains to grow at the sec14(ts) restrictive temperature. ... More
A non-exchangeable fluorescent phospholipid analog as a membrane traffic marker of the endocytic pathway.
AuthorsWillem J, ter Beest M, Scherphof G, Hoekstra D
JournalEur J Cell Biol
PubMed ID2076704
'The fluorescent phospholipid analog N-(lissamine rhodamine B sulfonyl)phosphatidylethanolamine (N-Rh-PE) was inserted into the plasma membrane of Baby hamster kidney cells at low temperature (2 degrees C). The mobility characteristics of the analog--as revealed by fluorescence photobleaching recovery--were very similar to those of membrane-inserted 1-acyl-2[6-[N-(7-nitro-2,1,3-benzoxadiazol-4-yl) amino]caproyl] phosphatidylcholine (C6-NBD-PC). Upon warming to ... More
Membrane-bound plasma platelet activating factor acetylhydrolase acts on substrate in the aqueous phase.
'Human plasma platelet activating factor acetylhydrolase (pPAF-AH) is a phospholipase A(2) that specifically hydrolyzes the sn-2 ester of platelet activating factor (PAF) and of phospholipids with oxidatively truncated sn-2 fatty acyl chains. pPAF-AH is bound to lipoproteins in vivo, and it binds essentially irreversibly to anionic and zwitterionic phospholipid vesicles ... More
Synthesis and properties of radioiodinated phospholipid analogues that spontaneously undergo vesicle-vesicle and vesicle-cell transfer.
AuthorsSchroit AJ, Madsen JW
JournalBiochemistry
PubMed ID6615789
'An efficient method for the synthesis and purification of a variety of iodinated phospholipid analogues is described. 1-Acyl-2-[[[3-(3-[125I]iodo-4-hydroxyphenyl)- propionyl]amino]caproyl]phosphatidylcholine (125I-PC) was prepared by alkylation of 1-acyl-2-(aminocaproyl)phosphatidylcholine with monoiodinated Bolton-Hunter reagent. 125I-Labeled phosphatidic acid, phosphatidylethanolamine, and phosphatidylserine were produced from 125I-PC by phospholipase D catalyzed base exchange in the presence of ... More
Phospholipid scramblase activation pathways in lymphocytes.
AuthorsWilliamson P, Christie A, Kohlin T, Schlegel RA, Comfurius P, Harmsma M, Zwaal RF, Bevers EM
JournalBiochemistry
PubMed ID11434775
'In erythrocytes and platelets, activation of a nonspecific lipid flipsite termed the scramblase allows rapid, bidirectional transbilayer movement of all types of phospholipids. When applied to lymphoid cells, scramblase assays reveal a similar activity, with scrambling rates intermediate between those seen in platelets and erythrocytes. Scrambling activity initiated in lymphoid ... More
Polar angle as a determinant of amphipathic alpha-helix-lipid interactions: a model peptide study.
AuthorsUematsu N, Matsuzaki K
JournalBiophys J
PubMed ID11023911
'Various physicochemical properties play important roles in the membrane activities of amphipathic antimicrobial peptides. To examine the effects of the polar angle, two model peptides, thetap100 and thetap180, with polar angles of 100 degrees and 180 degrees, respectively, were designed, and their interactions with membranes were investigated in detail. These ... More
Lipid effects on aggregation of pulmonary surfactant protein SP-C studied by fluorescence energy transfer.
AuthorsHorowitz AD, Baatz JE, Whitsett JA
JournalBiochemistry
PubMed ID8373758
'The self-association of pulmonary surfactant protein SP-C in lipid vesicles was studied using fluorescence energy transfer. Bovine SP-C was labeled with two fluorescent probes, succinimidyl 6-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]hexanoate and eosin isothiocyanate, on the amino terminus of the protein, producing NBD-SP-C and EITC-SP-C, respectively. The N-terminus of SP-C was relatively immobile between 20 ... More
Thermodynamics and kinetics of phospholipid monomer-vesicle interaction.
AuthorsNichols JW
JournalBiochemistry
PubMed ID4084528
'Resonance energy transfer between acyl chain labeled (7-nitro-2,1,3-benzoxadiazol-4-yl)phosphatidylcholine (NBD-PC) and head group labeled (lissamine rhodamine B sulfonyl)phosphatidylethanolamine (N-Rh-PE) was used to monitor the rate of NBD-PC transfer between two populations of dioleoylphosphatidylcholine (DOPC) vesicles. Equilibration of NBD-PC between DOPC vesicles occurs by the diffusion of soluble monomers through the water ... More
Mechanisms governing the level of susceptibility of erythrocyte membranes to secretory phospholipase A2.
AuthorsJensen LB, Burgess NK, Gonda DD, Spencer E, Wilson-Ashworth HA, Driscoll E, Vu MP, Fairbourn JL, Judd AM, Bell JD
JournalBiophys J
PubMed ID15681653
'Although cell membranes normally resist the hydrolytic action of secretory phospholipase A(2) (sPLA(2)), they become susceptible during apoptosis or after cellular trauma. Experimentally, susceptibility to the enzyme can be induced by loading cells with calcium. In human erythrocytes, the ability of the calcium ionophore to cause susceptibility depends on temperature, ... More
Insulin effect on translational diffusion of lipids and proteins in the plasma membrane of isolated rat hepatocytes.
AuthorsStuschke M, Bojar H
JournalBiochim Biophys Acta
PubMed ID3890961
'The effects of insulin (10(-10)-10(-8) mol/l) on lateral diffusion of three fluorescent lipid probes, 1-acyl-2-(N-4-nitrobenzo-2-oxa-1,3-diazole)aminocaproyl phosphatidylcholine (NBD-PC), 5-(N-hexadecanoyl)aminofluorescein (F-C16), 5-(N-dodecanoyl)aminofluorescein (F-C12), and of fluorescein isothiocyanate-labeled proteins in the plasma membrane of intact rat hepatocytes were studied by the technique of fluorescence recovery after photobleaching. The absolute lateral diffusion coefficients of ... More
Transbilayer movement of a fluorescent phosphatidylethanolamine analogue across the plasma membranes of cultured mammalian cells.
AuthorsSleight RG, Pagano RE
JournalJ Biol Chem
PubMed ID3968058
'The internalization of a fluorescent analogue of phosphatidylethanolamine following its insertion into the plasma membrane of cultured Chinese hamster fibroblasts was examined. When liposomes composed of 50 mol % 1-acyl-2-(N-4-nitrobenzo-2-oxa-1,3-diazole)-aminocaproyl phosphatidylethanolamine (C6-NBD-PE) and dioleoylphosphatidylcholine were incubated with monolayer cell cultures at 2 degrees C, a spontaneous transfer of the fluorescent ... More
Hydrolysis of a fluorescent phospholipid substrate by phospholipase A2 and lipoprotein lipase.
AuthorsWittenauer LA, Shirai K, Jackson RL, Johnson JD
JournalBiochem Biophys Res Commun
PubMed ID6704113
'The fluorescent phospholipid 1-acyl-2-[6-[(7-nitro-2,1,3benzoxadiazol-4 -yl) amino]-caproyl] phosphatidylcholine (C6-NBD-PC) was used as a substrate for porcine pancreatic phospholipase A2 (PA2) and bovine milk lipoprotein lipase (LpL). Hydrolysis of C6-NBD-PC by either enzyme resulted in a greater than 50-fold fluorescence enhancement with no shift in the emission maximum at 540 nm; Ca++ ... More
The End2 mutation in CHO cells slows the exit of transferrin receptors from the recycling compartment but bulk membrane recycling is unaffected.
AuthorsPresley JF, Mayor S, Dunn KW, Johnson LS, McGraw TE, Maxfield FR
JournalJ Cell Biol
PubMed ID8376460
'We have characterized a new CHO cell line (12-4) derived from a parental line, TRVb-1, that expresses the human transferrin receptor. This mutant belongs to the end2 complementation group of endocytosis mutants. Like other end2 mutants, the endosomes in 12-4 cells show a partial acidification defect. These cells internalize LDL ... More
Lateral diffusion of lipids in renal cells: effects of hypoxia and reoxygenation and role of cytoskeleton.
AuthorsPaller MS
JournalAm J Physiol
PubMed ID8430768
'The effects of hypoxia and reoxygenation on the lateral mobility of membrane lipids were studied in primary cultures of rat proximal tubule epithelial cells at a subconfluent stage. The lipid lateral diffusion coefficient (DL) of the fluorescent probe 1-acyl-2-(N-4-nitrobenzo-2-oxa-1,3-diazol)-aminocaproyl phosphatidylethanolamine was determined in membranes of attached cells by the technique ... More
Polyamine inhibition of transbilayer movement of plasma membrane phospholipids in the erythrocyte ghost.
AuthorsBratton DL
JournalJ Biol Chem
PubMed ID8077200
'The resting plasma membrane of circulating blood cells demonstrates an asymmetric distribution of the phospholipid classes across the bilayer that is altered during cellular activation. To better understand the mechanisms governing transbilayer distribution of phospholipids, studies were conducted using the erythrocyte ghost, in which plasma membrane leaflet distribution of phospholipids ... More
Analysis of phospholipid transfer during HDL binding to platelets using a fluorescent analog of phosphatidylcholine.
AuthorsMartin-Nizard F, Richard B, Torpier G, Nouvelot A, Fruchart JC, Duthilleul P, Delbart C
JournalThromb Res
PubMed ID2820079
'Electron microscopic examination of the interaction of gold labelled HDL with platelets indicates that the internalized lipoprotein becomes closely associated with surface connected canaliculae and endocytic vacuoles. At the same time granule centralization occurs. Using fluorescent derivatives of naturally occurring lipids we have further investigated lipid exchange between HDL and ... More
Vesicular and nonvesicular transport of phosphatidylcholine in polarized HepG2 cells.
AuthorsWüstner D, Mukherjee S, Maxfield FR, Müller P, Herrmann A
JournalTraffic
PubMed ID11285138
'We have investigated the transport and canalicular enrichment of fluorescent phosphatidylcholine (PC) in HepG2 cells using the fluorescent analogs of PC C6-NBD-PC and beta-BODIPY-PC. Fluorescent PC was efficiently transported to the biliary canaliculus (BC) and became enriched on the lumenal side of the canalicular membrane as shown for C6-NBD-PC. Some ... More
Salvage of glucosylceramide by recycling after internalization along the pathway of receptor-mediated endocytosis.
AuthorsKok JW, Eskelinen S, Hoekstra K, Hoekstra D
JournalProc Natl Acad Sci U S A
PubMed ID2690077
'To examine the (intra)cellular fate of a glycolipid, normally residing at the cell surface, a fluorescent analog of glucosylceramide, 6-[N-(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]hexanoylglucosylsp hingosine (C6-NBD-glucosylceramide), was inserted into the plasma membrane of baby hamster kidney cells at low temperature. Upon warming the cells to 37 degrees C, part of the glycolipid analog was ... More
Flow cytometric analysis of transmembrane phospholipid movement in bull sperm.
'Fluorescent phospholipids are useful to investigate phospholipid dynamics in biological membranes. We used flow cytometry to investigate transbilayer phospholipid movement in live sperm cells. Acyl-labeled N-4-nitrobenzo-2-oxa-1,3-diazole (NBD) -phosphatidylcholine (-PC), -phosphatidylethanolamine (-PE), or -phosphatidylserine (-PS) were incorporated into sperm cells, and the transbilayer location was determined by extraction of probe from ... More
Fluorescent probes for living cells.
AuthorsJohnson I
JournalHistochem J
PubMed ID10188922
'The functional characteristics of fluorescent probes used for imaging and measuring dynamic processes in living cells are reviewed. Initial consideration is given to general design requirements for delivery, targeting, detectability and fluorescence readout, and current technologies for attaining them. Discussion then proceeds to the more application-specific properties of intracellular ion ... More
Insertion of fluorescent phosphatidylserine into the plasma membrane of red blood cells. Recognition by autologous macrophages.
AuthorsTanaka Y, Schroit AJ
JournalJ Biol Chem
PubMed ID6885820
'The interaction of macrophages with red blood cells (RBC) displaying phosphatidylserine (PS) in their surface membranes was investigated after the transfer of an exogenously supplied fluorescent lipid analog to the RBC. Nonfluorescent (quenched) lipid vesicles were formed by ultrasonication from 1-acyl-2-[(N-4-nitro-benzo-2-oxa-1,3 diazole)aminocaproyl]phosphatidyl-serine (NBD-PS) or 1-acyl-2[(N-4-nitrobenzo-2-oxa-1,3 diazole)aminocaproyl]phosphatidylcholine (NBD-PC). The interaction of ... More
Investigation on lipid asymmetry using lipid probes: Comparison between spin-labeled lipids and fluorescent lipids.
AuthorsDevaux PF, Fellmann P, Hervé P
JournalChem Phys Lipids
PubMed ID12093538
'Synthetic lipids with a nitroxide or a fluorescent probe have been extensively used during the last 30 years to determine the transmembrane diffusion of phospholipids in artificial or biological membranes. However, the relevance of data obtained with these modified lipids has sometimes been questioned. Beside possible artefacts introduced by the ... More
Lipid domains and lipid/protein interactions in biological membranes.
AuthorsTocanne JF, Cézanne L, Lopez A, Piknova B, Schram V, Tournier JF, Welby M
JournalChem Phys Lipids
PubMed ID8001179
'In the fluid mosaic model of membranes, lipids are organized in the form of a bilayer supporting peripheral and integral proteins. This model considers the lipid bilayer as a two-dimensional fluid in which lipids and proteins are free to diffuse. As a direct consequence, both types of molecules would be ... More
Metabolism and intracellular distribution of a fluorescent analogue of phosphatidic acid in cultured fibroblasts.
AuthorsPagano RE
JournalAnn N Y Acad Sci
PubMed ID6584075
'We have shown that a fluorescent compound, C6-NBD-PA, behaves as an analogue for phosphatidic acid, an important intermediate in glycerolipid biosynthesis. This derivative is preferentially transferred from phospholipid vesicles to cultured Chinese hamster fibroblasts at 2 degrees C, while the C6-NBD-PA-derived fluorescence is localized at the nuclear membrane, endoplasmic reticulum, ... More
Identification and characterization of a gene encoding phospholipase D activity in yeast.
AuthorsWaksman M, Eli Y, Liscovitch M, Gerst JE
JournalJ Biol Chem
PubMed ID8576189
'We have identified an open reading frame on chromosome XI of the yeast, Saccharomyces cerevisiae, as encoding a protein with phospholipase D (PLD) activity. We have named this open reading frame, PLD1, and show that yeast bearing a disruption in this gene are unable to catalyze the hydrolysis of phosphatidylcholine. ... More
The lantibiotic nisin induces transmembrane movement of a fluorescent phospholipid.
AuthorsMoll GN, Konings WN, Driessen AJ
JournalJ Bacteriol
PubMed ID9852000
'Nisin is a pore-forming antimicrobial peptide. The capacity of nisin to induce transmembrane movement of a fluorescent phospholipid in lipid vesicles was investigated. Unilamellar phospholipid vesicles that contained a fluorescent phospholipid (1-acyl-2-(6-[(7-nitro-2-1, 3-benzoxadiazol-4-yl)amino]caproyl)-sn-glycero-3-phosphocholine) in the inner leaflet of the bilayer were used. Nisin-induced movement of the fluorescent phospholipid from the ... More
Determination of the transbilayer distribution of fluorescent lipid analogues by nonradiative fluorescence resonance energy transfer.
AuthorsWolf DE, Winiski AP, Ting AE, Bocian KM, Pagano RE
JournalBiochemistry
PubMed ID1550813
'We measured the nonradiative fluorescence resonance energy transfer between 7-nitro-2,1,3-benzoxadiazol-4-yl (NBD) labeled lipids (amine labeled phosphatidylethanolamine or acyl chain labeled phosphatidylcholine) and rhodamine labeled lipids in large unilamellar dioleoylphosphatidylcholine vesicles. Two new rhodamine labeled lipid analogues, one a derivative of monolauroylphosphatidylethanolamine and the other of sphingosylphosphorylcholine, were found to exchange ... More
Transmembrane diffusion of fluorescent phospholipids in human erythrocytes.
AuthorsColleau M, Hervé P, Fellmann P, Devaux PF
JournalChem Phys Lipids
PubMed ID2060062
'The outside-inside passage and transmembrane equilibrium distribution of several amphiphilic fluorescent phospholipids were examined in human erythrocytes. The results were compared with previous kinetic data obtained with spin-labeled phospholipids and with the equilibrium distribution of endogenous lipids in erythrocytes. When a nitro benzoxadiazole (NBD) was at the terminal position of ... More
Phospholipid flop induced by transmembrane peptides in model membranes is modulated by lipid composition.
AuthorsKol MA, van Laak AN, Rijkers DT, Killian JA, de Kroon AI, de Kruijff B
JournalBiochemistry
PubMed ID12515559
'Since phospholipid synthesis is generally confined to one leaflet of a membrane, membrane growth requires phospholipid translocation (flip-flop). It is generally assumed that this process is protein-mediated; however, the mechanism of flip-flop remains elusive. Previously, we have demonstrated flop of 2-[6-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]caproyl] (C6NBD) phospholipids, induced by the presence of membrane-spanning peptides ... More
Determination of lipid asymmetry in human red cells by resonance energy transfer.
AuthorsConnor J, Schroit AJ
JournalBiochemistry
PubMed ID3663645
'This report describes the application of a resonance energy transfer assay to determine the transbilayer distribution of 7-nitro-2,1,3-benzoxadiazol-4-yl (NBD)-labeled lipid analogues. The validity of this technique was established by determining the relationship between the distance of separation of lissamine rhodamine B labeled phosphatidylethanolamine (N-Rho-PE) acceptor lipid and NBD-labeled donor lipid ... More
Reversed-phase liquid chromatographic method for the determination of 7-nitrobenz-2-oxa-1,3-diazol-4-yl-labelled lipid analogues.
AuthorsFrijters CM, Tuijn CJ, Hoek FJ, Groen AK, Oude Elferink RP, Zegers BN
JournalJ Chromatogr B Biomed Sci Appl
PubMed ID9686866
'This paper reports the development of a dual column system for the simultaneous separation of fluorescent short-chain ceramide, 6-[(7-nitrobenz-2-oxa-1,3,-diazol-4-yl[NBD])amino]hexanoyl-sphingo sine and its metabolites, C6-NBD-sphingomyelin and C6-NBD-glucosylceramide, as well as the fluorescent derivatives of choline and serine phosphatides. The method enables the separation of these lipids in a single run on ... More
Translocation of fluorescent ether phospholipid, but not its diacyl counterpart, after insertion in plasma membranes of control and plasmalogen-deficient fibroblasts.
AuthorsVan der Veer E, Van der Weide D, Heijmans HS, Hoekstra D
JournalBiochim Biophys Acta
PubMed ID8452863
'Fluorescently labelled ether phospholipid (1-O-alkyl/alkenyl-2-acyl- glycerophosphocholine) readily internalizes at low temperatures (2 degrees C) after insertion into the plasma membrane of cultured fibroblasts. This fate differs markedly from that of its diacyl phospholipid analogue, which remains associated with the plasma membrane under similar conditions. Analysis by thin-layer chromatography reveals that ... More
Phosphatidylinositol 4,5-bisphosphate synthesis is required for activation of phospholipase D in U937 cells.
AuthorsPertile P, Liscovitch M, Chalifa V, Cantley LC
JournalJ Biol Chem
PubMed ID7890622
'Phospholipase D (PLD) has been implicated in signal transduction and membrane traffic. We have previously shown that phosphatidylinositol 4,5-bisphosphate (PtdIns-4,5-P2) stimulates in vitro partially purified brain membrane PLD activity, defining a novel function of PtdIns-4,5-P2 as a PLD cofactor. In the present study we extend these observations to permeabilized U937 ... More
Membrane fluidity in normal and cystic fibrosis fibroblasts.
AuthorsChristian ST, Monti JA, Finley WH
JournalBiochem Biophys Res Commun
PubMed ID597319
Differential rotational mobility of a membrane-bound fluorochrome in cell types of increasing oncogenic potential.
AuthorsMonti JA, Sarrif AM, Christian ST, Saxholm HJ
JournalArch Biochem Biophys
PubMed ID111623
Synthesis and properties of a fluorescent derivative of phosphatidylcholine.
AuthorsMonti JA, Christian ST, Shaw WA, Finley WH
JournalLife Sci
PubMed ID895370
Changes in the organization of the sea urchin egg plasma membrane upon fertilization: indications from the lateral diffusion rates of lipid-soluble fluorescent dyes.
AuthorsWolf DE, Kinsey W, Lennarz W, Edidin M
JournalDev Biol
PubMed ID7461282
Resonance energy transfer assay of protein-mediated lipid transfer between vesicles.
AuthorsNichols JW, Pagano RE
JournalJ Biol Chem
PubMed ID6853520
Diffusion of low density lipoprotein-receptor complex on human fibroblasts.
AuthorsBarak LS, Webb WW
JournalJ Cell Biol
PubMed ID6296157
Lateral diffusion of lipids in model and natural membranes.
AuthorsTocanne JF, Dupou-Cézanne L, Lopez A
JournalProg Lipid Res
PubMed ID8022844
Comparative lateral diffusion of fluorescent lipid analogues in phospholipid multibilayers.
AuthorsDerzko Z, Jacobson K
JournalBiochemistry
PubMed ID7470449
Synthesis of a fluorescent analogue of phosphatidylcholine.
AuthorsSchmidt N, Gercken G
JournalChem Phys Lipids
PubMed ID4085088
A fluorescent analogue of phosphatidylcholine was synthesized by acylation of 1-oleoyl-sn-glycero-3-phosphocholine with 6-N-(tert-butyloxycarbonyl)aminocaproic acid anhydride employing the catalyst 4-pyrrolidinopyridine. Removal of the protective group by treatment with HCl in chloroform was followed by subsequent reaction with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) to form the fluorescent analogue of phosphatidylcholine, 1-oleoyl-2-(NBD)aminocaproyl-sn-glycero-3-phosphocholine, in good yield and ... 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
A subfamily of P-type ATPases with aminophospholipid transporting activity.
AuthorsTang X, Halleck MS, Schlegel RA, Williamson P
JournalScience
PubMed ID8633245
The appearance of phosphatidylserine on the surface of animal cells triggers phagocytosis and blood coagulation. Normally, phosphatidylserine is confined to the inner leaflet of the plasma membrane by an aminophospholipid translocase, which has now been cloned and sequenced. The bovine enzyme is a member of a previously unrecognized subfamily of ... More
A facile method for direct determination of phospholipase A2 activity in intact cells.
AuthorsDagan A, Yedgar S
JournalBiochem Int
PubMed ID3435543
The fluorescent phospholipid analogue 1-acyl-2-(N-4-nitrobenzo-2-oxa-1,3- diazole)aminocaproylphosphatidylcholine (C6-NBD-PC), which incorporates into cell membranes, is employed as a substrate for phospholipase A2 (PLA2) in intact cells. The interaction of this substrate with the cells produces only one fluorescent product; the fatty acid C6-NBD-FA, which does not incorporate into other lipids, and is ... More
Two-dimensional crystals of streptavidin on biotinylated lipid layers and their interactions with biotinylated macromolecules.
AuthorsDarst SA, Ahlers M, Meller PH, Kubalek EW, Blankenburg R, Ribi HO, Ringsdorf H, Kornberg RD
JournalBiophys J
PubMed ID1901232
Streptavidin forms two-dimensional crystals when specifically bound to layers of biotinylated lipids at the air/water interface. The three-dimensional structure of streptavidin determined from the crystals by electron crystallography corresponds well with the structure determined by x-ray crystallography. Comparison of the electron and x-ray crystallographic structures reveals the occurrence of free ... More
Islet amyloid polypeptide-induced membrane leakage involves uptake of lipids by forming amyloid fibers.
AuthorsSparr E, Engel MF, Sakharov DV, Sprong M, Jacobs J, de Kruijff B, Höppener JW, Killian JA
JournalFEBS Lett
PubMed ID15527771
Fibril formation of islet amyloid polypeptide (IAPP) is associated with cell death of the insulin-producing pancreatic beta-cells in patients with Type 2 Diabetes Mellitus. A likely cause for the cytotoxicity of human IAPP is that it destroys the barrier properties of the cell membrane. Here, we show by fluorescence confocal ... More
Use of a fluorescence assay to monitor the kinetics of fusion between erythrocyte ghosts, as induced by Sendai virus.
AuthorsHoekstra D, Klappe K
JournalBiosci Rep
PubMed ID3034350
The kinetics of the fusion process between erythrocyte ghosts, as induced by Sendai virus, were readily revealed by a simple fluorescence procedure previously employed to characterize the fusion of viruses with biological membranes. The method relies on the relief of fluorescence selfquenching of the membrane-inserted probe octadecyl Rhodamine B chloride ... More
Kinetics of soluble lipid monomer diffusion between vesicles.
AuthorsNichols JW, Pagano RE
JournalBiochemistry
PubMed ID7248248
The fluorescent phospholipid 1-acyl-2-[12-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]dodecanoyl]phosphatidylcholine (C12-NBD-PC) was used to study the kinetics of lipid transfer between phospholipid vesicles. A model based on lipid transfer resulting from the diffusion of soluble monomers was found to accurately predict the kinetics of this transfer process. From these studies, we conclude that (i) C12-NBD-PC transfer ... More
Miscibility of lipoteichoic acid in dipalmitoylphosphatidylcholine studied by monofilm investigations and fluorescence microscopy.
AuthorsGutberlet T, Milde K, Bradaczek H, Haas H, Möhwald H
JournalChem Phys Lipids
PubMed ID8181104
The miscibility of the bacterial amphiphile lipoteichoic acid, a constituent of the cytoplasmic membrane of Gram-positive bacteria, in dipalmitoylphosphatidylcholine has been investigated by classic monofilm measurements and fluorescence microscopy at the air-water interface of monofilms obtained by spreading mixtures of both amphiphiles on a water subphase. The isotherms indicated miscibility ... More
High-throughput fluorescence assay for small-molecule inhibitors of autophagins/atg4.
Autophagy is an evolutionarily conserved process for catabolizing damaged proteins and organelles in a lysosome-dependent manner. Dysregulation of autophagy may cause various diseases, such as cancer and neurodegeneration. However, the relevance of autophagy to diseases remains controversial because of the limited availability of chemical modulators. Herein, the authors developed a ... More
Relationship between membrane physical properties and secretory phospholipase A2 hydrolysis kinetics in S49 cells during ionophore-induced apoptosis.
AuthorsBailey RW, Olson ED, Vu MP, Brueseke TJ, Robertson L, Christensen RE, Parker KH, Judd AM, Bell JD,
JournalBiophys J
PubMed ID17545239
During apoptosis, changes occur in lymphocyte membranes that render them susceptible to hydrolysis by secretory phospholipase A(2) (sPLA(2)). To study the relevant mechanisms, a simplified model of apoptosis using a calcium ionophore was applied. Kinetic and flow cytometry experiments provided key observations regarding ionophore treatment: the initial rate of hydrolysis ... More
A novel membrane protein, Ros3p, is required for phospholipid translocation across the plasma membrane in Saccharomyces cerevisiae.
AuthorsKato U, Emoto K, Fredriksson C, Nakamura H, Ohta A, Kobayashi T, Murakami-Murofushi K, Kobayashi T, Umeda M,
JournalJ Biol Chem
PubMed ID12133835
Ro09-0198 (Ro) is a tetracyclic peptide antibiotic that binds specifically to phosphatidylethanolamine (PE) and causes cytolysis. To investigate the molecular basis of transbilayer movement of PE in biological membranes, we have isolated a series of budding yeast mutants that are hypersensitive to the Ro peptide. One of the most sensitive ... More
Interactions of histone H1 with phospholipids and comparison of its binding to giant liposomes and human leukemic T cells.
AuthorsZhao H, Bose S, Tuominen EK, Kinnunen PK,
JournalBiochemistry
PubMed ID15287747
Due to its net positive charge histone H1 readily associates with liposomes containing acidic phospholipids, such as phosphatidylserine (PS). Interestingly, circular dichroism reveals that while histone H1 in aqueous solutions appears as a random coil, its binding to liposomes containing PS is associated with a pronounced increase in alpha-helicity and ... More
Synthesis of fluorescent and radiolabeled analogues of phosphatidic acid.
AuthorsLongmuir KJ, Martin OC, Pagano RE
JournalChem Phys Lipids
PubMed ID4006033
Procedures for the synthesis of fluorescent and radiolabeled analogues of phosphatidic acid are described. The fluorophore 7-nitrobenzo-2-oxa-1,3-diazole (NBD) was coupled to 6-amino-caproic acid and 12-aminododecanoic acid by reaction of NBD-chloride with the amino acids under mild alkaline conditions at room temperature. 1,2-Dioleoyl-sn-[U-14C]glycerol 3-phosphate was prepared by acylation of sn-[U-14C]glycerol 3-phosphate ... More
Interactions of adriamycin, cytochrome c, and serum albumin with lipid monolayers containing poly(ethylene glycol)-ceramide.
Poly(ethylene glycol)(2000)C(20)ceramide (PEG-Cer) containing monolayers at an air/water interface were characterized by measuring their surface pressure versus area/molecule (pi-A) and surface potential versus area/molecule (Delta V-A) isotherms. The behavior of pi-A as well as Delta V versus lipid density (Delta V-n) and Delta V-pi isotherms for PEG-Cer are in keeping ... More
Lateral diffusion of specific antibodies bound to lipid monolayers on alkylated substrates.
AuthorsSubramaniam S, Seul M, McConnell HM
JournalProc Natl Acad Sci U S A
PubMed ID3006037
We have measured the lateral mobility of fluoresceinated monoclonal IgG antibodies bound specifically to a spin label lipid hapten in phospholipid monolayers supported on alkylated silicon oxide surfaces. Dimyristoyl phosphatidylcholine and dipalmitoyl phosphatidylcholine monolayers containing 5 mol% of the lipid hapten were transferred by conventional Langmuir-Blodgett techniques onto substrates alkylated ... More
Topographical distribution of a membrane-inserted fluorescent phospholipid analogue during cell fusion.
AuthorsHoekstra D
JournalExp Cell Res
PubMed ID6301868
Potential alterations in the transbilayer distribution of lipid molecules during cell-cell fusion were studied, using the fluorescent phospholipid analogue 1-acyl, 2-(N-4-nitrobenzo-2-oxa-1,3-diazole)-aminocaproyl phosphatidylcholine (C6-NBD-PC). The fluophore was inserted into the outer leaflet of the plasma membrane of Chinese hamster fibroblasts from an exogenous source and cell-cell fusion was induced either with ... More
Two-dimensional chiral crystals of phospholipid.
AuthorsWeis RM, McConnell HM
JournalNature
PubMed ID6738702
Epifluorescence optical microscopy has been used to show the formation of solid phase domains from fluid phase domains on compression of dipalmitoylphosphatidylcholine (DPPC) monolayers at the air-water interface. These monolayers can be transferred to solid substrates, and in certain conditions exhibit X-ray diffraction characteristic of highly ordered two-dimensional crystals. In ... More
The Thy-1 antigen exhibits rapid lateral diffusion in the plasma membrane of rodent lymphoid cells and fibroblasts.
AuthorsIshihara A, Hou Y, Jacobson K
JournalProc Natl Acad Sci U S A
PubMed ID2881297
Thy-1 is a plasma membrane protein, but its primary structure lacks the typical membrane-spanning sequence. Recent studies revealed that a glycophospholipid is covalently bound to the carboxyl terminus, suggesting that the protein is integrated into the plasma membrane by this lipid moiety. Lateral diffusion of Thy-1 was measured in mouse ... More
Rapid determination of the transbilayer distribution of NBD-phospholipids in erythrocyte membranes with dithionite.
AuthorsPomorski T, Herrmann A, Zachowski A, Devaux PF, Müller P
JournalMol Membr Biol
PubMed ID8019600
The assessment of the transverse distribution and mobility of NBD-labelled phospholipid analogues in biological membranes by selective chemical destruction of fluorescent label in the outer monolayer with dithionite has been investigated using resealed erythrocyte ghosts as a model system. The distribution of those analogues can be determined in < 30 ... More
The Fluorosome technique for investigating membrane on- and off-loading of drugs by beta-CD and sonicated SUV.
AuthorsFix M, Melchior DL
JournalFEBS Lett
PubMed ID11959114
The application of the Fluorosome technique to test drug delivery systems is described. Fluorosomes, egg phosphatidylcholine liposomes with bilayer embedded fluorophores, were employed to investigate the ability of sonicated small unilamellar vesicles (sSUV) and beta-cyclodextrins (beta-CD) to deliver drugs into or extract drugs from the fluorosome's phospholipid bilayer. The addition ... More
Regulation of phospholipid scramblase activity during apoptosis and cell activation by protein kinase Cdelta.
AuthorsFrasch SC, Henson PM, Kailey JM, Richter DA, Janes MS, Fadok VA, Bratton DL
JournalJ Biol Chem
PubMed ID10770950
Phospholipid scramblase induces nonspecific bidirectional movement of phospholipids across the membrane during cell activation and has been proposed to mediate the appearance of phosphatidylserine (PS) in the plasma membrane outer leaflet during apoptosis, a cell surface change that is critical for apoptotic cell removal. We report here that protein kinase ... More
Chemistry and biology of N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-labeled lipids: fluorescent probes of biological and model membranes.
AuthorsChattopadhyay A
JournalChem Phys Lipids
PubMed ID2191793
Lipids that are covalently labeled with the 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD) group are widely used as fluorescent analogues of native lipids in model and biological membranes to study a variety of processes. The fluorescent NBD group may be attached either to the polar or the apolar regions of a wide variety of ... More
Large-scale preparation of asymmetrically labeled fluorescent lipid vesicles.
AuthorsDao HN, McIntyre JC, Sleight RG
JournalAnal Biochem
PubMed ID1888035
A method for producing lipid vesicles containing fluorescent phospholipid analogues localized to the inner leaflet of their membrane was developed. Incubation of a 450-fold molar excess of serum albumin with lipid vesicles symmetrically labeled with 1 mol % 1-palmitoyl-2-(N-4-nitrobenzo-2-oxa-1,3-diazolyl)amino-caproyl phosphatidylcholine resulted in the removal of 99% of the fluorescent lipid ... More
Characterization of lipid domains in erythrocyte membranes.
AuthorsRodgers W, Glaser M
JournalProc Natl Acad Sci U S A
PubMed ID1996337
Fluorescence digital imaging microscopy was used to study the lateral distribution of the lipid components in erythrocyte membranes. Intact erythrocytes labeled with phospholipids containing a fluorophore attached to one fatty acid chain showed an uneven distribution of the phospholipids in the membrane thereby demonstrating the presence of membrane domains. The ... More
Transbilayer transport of ions and lipids coupled with mastoparan X translocation.
AuthorsMatsuzaki K, Yoneyama S, Murase O, Miyajima K
JournalBiochemistry
PubMed ID8679603
The transbilayer movement of ions and lipids induced by mastoparan X, a peptidic toxin from Vespa xanthoptera, was investigated by use of lipid vesicles as a model membrane system. Negatively charged phosphatidylglycerol remarkably enhanced the peptide-lipid interactions. Mastoparan X induced the ion flow by forming a short-lived, multimeric pore in ... More
Human cytomegalovirus carries a cell-derived phospholipase A2 required for infectivity.
AuthorsAllal C, Buisson-Brenac C, Marion V, Claudel-Renard C, Faraut T, Dal Monte P, Streblow D, Record M, Davignon JL
JournalJ Virol
PubMed ID15220446
Human cytomegalovirus (HCMV) is known to carry host cell-derived proteins and mRNAs whose role in cell infection is not understood. We have identified a phospholipase A2 (PLA2) activity borne by HCMV by using an assay based on the hydrolysis of fluorescent phosphatidylcholine. This activity was found in all virus strains ... More
Effects of lung surfactant proteolipid SP-C on the organization of model membrane lipids: a fluorescence study.
AuthorsHorowitz AD, Elledge B, Whitsett JA, Baatz JE
JournalBiochim Biophys Acta
PubMed ID1616924
Lipid-protein interactions of pulmonary surfactant-associated protein SP-C in model DPPC/DPPG and DPPC/DPPG/eggPC vesicles were studied using steady-state and time-resolved fluorescence measurements of two fluorescent phospholipid probes, NBD-PC and NBD-PG. These fluorescent probes were utilized to determine SP-C-induced lipid perturbations near the bilayer surface, and to investigate possible lipid headgroup-specific interactions ... More