Use of resonance energy transfer to study the kinetics of amphiphile transfer between vesicles.
AuthorsNichols JW, Pagano RE
JournalBiochemistry
PubMed ID7082641
Effects of a cationic and hydrophobic peptide, KL4, on model lung surfactant lipid monolayers.
AuthorsMa J, Koppenol S, Yu H, Zografi G
JournalBiophys J
PubMed ID9545051
'We report on the surface behavior of a hydrophobic, cationic peptide, [lysine-(leucine)4]4-lysine (KL4), spread at the air/water interface at 25 degrees C and pH 7.2, and its effect at very low molar ratios on the surface properties of the zwitterionic phospholipid 1,2-dipalmitoylphosphatidylcholine (DPPC), and the anionic forms of 1-palmitoyl-2-oleoylphosphatidylglycerol (POPG) ... More
Low concentrations of bile salts increase the rate of spontaneous phospholipid transfer between vesicles.
AuthorsNichols JW
JournalBiochemistry
PubMed ID3768300
'The rate of 1-palmitoyl-2-[12-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino] dodecanoyl] phosphatidylcholine (P-C12-NBD-PC) transfer between dioleoylphosphatidylcholine vesicles was measured by a technique based on resonance energy transfer between P-C12-NBD-PC and N-(lissamine rhodamine B sulfonyl)dioleoylphosphatidylethanolamine [Nichols, J. W., & Pagano, R. E. (1982) Biochemistry 21, 1720-1726]. Addition of bile salts at concentrations below their critical micelle concentrations ... More
Role of surface lysine residues of adipocyte fatty acid-binding protein in fatty acid transfer to phospholipid vesicles.
AuthorsLiou HL, Storch J
JournalBiochemistry
PubMed ID11371211
'The tertiary structure of murine adipocyte fatty acid-binding protein (AFABP) is a flattened 10-stranded beta-barrel capped by a helix-turn-helix segment. This helical domain is hypothesized to behave as a "lid" or portal for ligand entry into and exit from the binding cavity. Previously, we demonstrated that anthroyloxy-labeled fatty acid (AOFA) ... More
Kinetics of fluorescent-labeled phosphatidylcholine transfer between nonspecific lipid transfer protein and phospholipid vesicles.
AuthorsNichols JW
JournalBiochemistry
PubMed ID3378037
'Recently, rat liver nonspecific lipid transfer protein (nsLTP) was shown to form a fluorescent complex when allowed to equilibrate with self-quenching vesicles prepared from the fluorescent phospholipid 1-palmitoyl-2-[12-[(7-nitro-2,1,3-benzoxadiazol-4- yl)amino]dodecanoyl]phosphatidylcholine (P-C12-NBD-PC) [Nichols, J. W. (1987) J. Biol. Chem. 262, 14172-14177]. Investigation of the mechanism of complex formation was continued by studying ... More
Lipid binding to amyloid beta-peptide aggregates: preferential binding of cholesterol as compared with phosphatidylcholine and fatty acids.
AuthorsAvdulov NA, Chochina SV, Igbavboa U, Warden CS, Vassiliev AV, Wood WG
JournalJ Neurochem
PubMed ID9326304
'Amyloid beta-peptide (A beta) aggregates are one of the key neuropathological characteristics of Alzheimer''s disease. A beta belongs to a group of proteins that aggregate and form beta-sheets, and some of these proteins bind cholesterol and other lipids. The purpose of the experiments reported here was to determine if cholesterol, ... More
Lateral diffusion of erythrocyte phospholipids in model membranes comparison between inner and outer leaflet components.
AuthorsCribier S, Morrot G, Neumann JM, Devaux PF
JournalEur Biophys J
PubMed ID2155112
'The physical properties of lipid bilayers with a similar composition to the outer and inner leaflets of the human erythrocyte membrane have been examined in protein-free model systems. The outer leaflet (OL) was represented by a phospholipid mixture containing phosphatidylcholine and sphingomyelin extracted from human erythrocytes, while a mixture of ... More
Effect of lateral mobility of fluorescent probes in lipid mixing assays of cell fusion.
AuthorsHuang SK, Cheng M, Hui SW
JournalBiophys J
PubMed ID2291938
'Monolayers of human erythrocytes, immobilized on a cover slip, were induced to fuse by polyethylene glycol (mol wt 8,000). The mobility of fluorescent probes, 1-oleoyl-2-[12-[(7-nitro-2,1,3-benzoxadizol-4-yl)amino]dodecanoyl] phosphatidyl-choline (C12-NBD-PC), from labeled cells to unlabeled cells was monitored by video-enhanced fluorescence microscopy. A dequenching curve was obtained from the measurement of fluorescence intensities ... More
Phospholipid membranes form specific nonbilayer molecular arrangements that are antigenic.
AuthorsAguilar L, Ortega-Pierres G, Campos B, Fonseca R, Ibáñez M, Wong C, Farfán N, Naciff JM, Kaetzel MA, Dedman JR, Baeza I
JournalJ Biol Chem
PubMed ID10464237
'Hexagonal phase (H(II))-preferring lipids such as phosphatidate, cardiolipin, and phosphatidylserine form nonbilayer molecular arrangements in lipid bilayers. While their presence in biological membranes has not been established, in vitro studies suggest that alterations in membrane properties modify their function. In this study, antiphospholipid monoclonal antibodies were developed against nonbilayer structures. ... More
7-nitrobenz-2-oxa-1,3-diazole-4-yl-labeled phospholipids in lipid membranes: differences in fluorescence behavior.
AuthorsMazères S, Schram V, Tocanne JF, Lopez A
JournalBiophys J
PubMed ID8804615
'Steady-state and time-resolved fluorescence properties of the 7-nitrobenz-2-oxa-1, 3-diazole-4-yl (NBD) fluorophore attached either to the sn-2 acyl chain of various phospholipids (phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidic acid) or to the polar headgroup of phosphatidylethanolamine were studied after insertion of these NBD-labeled lipid probes into unilamellar vesicles of phosphatidylcholine, phosphatidylglycerol, phosphatidic ... More
Functional reconstitution of the olfactory membrane: incorporation of the olfactory adenylate cyclase in liposomes.
AuthorsAnholt RR
JournalBiochemistry
PubMed ID3064810
'Chemosensory cilia isolated from the olfactory epithelium of Rana catesbeiana were solubilized with Lubrol PX in the presence of supplementary lipid, forskolin, and sodium fluoride. Subsequent removal of the detergent by adsorption onto Biobeads SM2 results in the formation of proteoliposomes that display forskolin- and GTP gamma S-sensitive adenylate cyclase ... 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
Real-time fluorescence polarization measurements: interaction of phospholipase A2 with a fluorescent lecithin derivative.
AuthorsMonti JA, McBride MR, Barker SA, Linton JL, Christian ST
JournalJ Biochem Biophys Methods
PubMed ID3924989
'We have modified an SLM 4800 fluorometer to allow continuous measurement of fluorescence polarization. The modifications included construction of simple mechanical and electronic accessories which allowed polarizer position to be program-controlled by an HP 9815 calculator. With these modifications we were able to follow the interaction of the fluorescent lipid ... 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
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
Binding of fluorescent-labeled phosphatidylcholine to rat liver nonspecific lipid transfer protein.
AuthorsNichols JW
JournalJ Biol Chem
PubMed ID3654657
'The fluorescent-labeled phospholipid, 1-palmitoyl-2-[12-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]dodecan oyl] phosphatidylcholine (P-C12-NBD-PC), is highly self-quenched when contained as a high mole percent in phospholipid vesicles. The addition of nonspecific lipid transfer protein (nsLTP), which is identical to sterol carrier protein2, to P-C12-NBD-PC self-quenching vesicles results in a large increase in the fluorescence yield. Evidence is ... More
Raft-like domain formation in large unilamellar vesicles probed by the fluorescent phospholipid analogue, C12NBD-PC.
AuthorsCoste V, Puff N, Lockau D, Quinn PJ, Angelova MI
JournalBiochim Biophys Acta
PubMed ID16626630
'The liquid-ordered/disordered-phase domain co-existence in large unilamellar vesicle membranes consisting of phosphatidylcholine:sphingomyelin (2:1) with different amounts of cholesterol has been examined using a concentration-dependent self-quenching of a single reporter molecule, C12NBD-PC. A temperature-dependent decrease of fluorescence intensity was associated with the expected formation and increase of l(o)-phase membrane fraction in ... More
Phosphatidylserine receptors: role of CD36 in binding of anionic phospholipid vesicles to monocytic cells.
AuthorsTait JF, Smith C
JournalJ Biol Chem
PubMed ID9915844
'Exposure of phosphatidylserine (PtdSer) has been implicated in the recognition and phagocytosis of senescent and apoptotic cells, and CD36 has been proposed as one receptor protein that recognizes PtdSer and other anionic phospholipids. We investigated the binding of phospholipid vesicles to the monocytic leukemia cell lines THP-1 and J774A.1 with ... More
In vivo recognition and clearance of red blood cells containing phosphatidylserine in their plasma membranes.
AuthorsSchroit AJ, Madsen JW, Tanaka Y
JournalJ Biol Chem
PubMed ID3988747
'We have previously investigated the interaction of macrophages with red blood cells (RBC) displaying phosphatidylserine (PS) in their surface membranes after the transfer of the fluorescent lipid analog 1-acyl-2-[(N-4-nitrobenzo-2-oxa-1,3-diazole)aminocaproyl] phosphatidylserine to the RBC (Tanaka, Y., and Schroit, A. J. (1983) J. Biol. Chem. 258, 11335-11343). This derivative, which is rapidly ... More
Annexin IV reduces the rate of lateral lipid diffusion and changes the fluid phase structure of the lipid bilayer when it binds to negatively charged membranes in the presence of calcium.
AuthorsGilmanshin R, Creutz CE, Tamm LK
JournalBiochemistry
PubMed ID8031756
'Bovine annexin IV (endonexin) was bound to supported planar bilayers composed of 1-palmitoyl-2-oleoylphosphatidylcholine (POPC) in the first monolayer facing the substrate, and varying mole fractions of POPC, 1-palmitoyl-2-oleoylphosphatidylglycerol (POPG) and small amounts of the fluorescent lipid analogs NBD-PC or NBD-PG in the second monolayer facing the large aqueous compartment. Lateral ... More
Organization and dynamics of the proteolipid complexes formed by annexin V and lipids in planar supported lipid bilayers.
AuthorsCézanne L, Lopez A, Loste F, Parnaud G, Saurel O, Demange P, Tocanne JF
JournalBiochemistry
PubMed ID10052949
'The consequences of the binding of annexin V on its lateral mobility and that of lipids were investigated by means of experimental and simulated FRAP experiments. Experiments were carried out on planar supported bilayers (PC/PS 9:1 mol/mol mixtures) in the presence of 1 mM CaCl2 in the subphase. The probes ... More
The phospholipid flippase activity of gastric vesicles.
AuthorsSuzuki H, Kamakura M, Morii M, Takeguchi N
JournalJ Biol Chem
PubMed ID9099684
'We found that isolated gastric vesicles contain a novel Mg2+-ATP-dependent phospholipid translocation (flippase) activity. Fluorescence analogue of phosphatidylcholine, 2-(12-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl)amino)dodecanoyl-1-hexadecanoyl-sn-glycero-3- phosphocholine, was ATP-dependently translocated from the outer (cytosolic) to inner (luminal) leaflet of the lipid membrane bilayer of hog gastric vesicles. The translocation was saturable and depended on time and ... More
Calcium-dependent annexin V binding to phospholipids: stoichiometry, specificity, and the role of negative charge.
AuthorsMeers P, Mealy T
JournalBiochemistry
PubMed ID8218240
'Annexin V is a Ca(2+)-dependent, phospholipid-binding protein that may have one or more membrane-related functions. The binding of annexin V to phospholipids in a detergent micelle matrix was studied to attempt to determine directly the stoichiometry of specific phospholipid-binding sites and the importance of negative charge. When annexin V binds ... More
The signal sequence moves through a ribosomal tunnel into a noncytoplasmic aqueous environment at the ER membrane early in translocation.
AuthorsCrowley KS, Reinhart GD, Johnson AE
JournalCell
PubMed ID8513496
'The signal sequence is in an aqueous milieu at an early stage in the translocation of a nascent secretory protein across the endoplasmic reticulum membrane. This was determined using fluorescent probes incorporated into the signal sequence of fully assembled ribosome-nascent chain-membrane complexes: the fluorescence lifetimes revealed that the probes were ... 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
Interactions of pulmonary surfactant protein A with phospholipid monolayers change with pH.
AuthorsRuano ML, Nag K, Casals C, Pérez-Gil J, Keough KM
JournalBiophys J
PubMed ID10465757
'The interaction of pulmonary surfactant protein A (SP-A) labeled with Texas Red (TR-SP-A) with monolayers containing zwitterionic and acidic phospholipids has been studied at pH 7.4 and 4.5 using epifluorescence microscopy. At pH 7.4, TR-SP-A expanded the pi-A isotherms of film of dipalmitoylphosphatidylcholine (DPPC). It interacted at high concentration at ... 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
Cardiolipin binding a light chain from lupus-prone mice.
AuthorsPereira B, Benedict CR, Le A, Shapiro SS, Thiagarajan P
JournalBiochemistry
PubMed ID9477972
'Autoantibodies in systemic lupus erythematosus react with multiple epitopes on highly conserved molecules such as nucleic acids, cytoskeletal proteins, phospholipids, and phospholipid-binding proteins. Analysis of the heavy- and light-chain variable sequences (VH and VL) has shown that a restricted set of V genes gives rise to these autoantibodies. Several monoclonal ... More
Kinetics and thermodynamics of calcium-induced lateral phase separations in phosphatidic acid containing bilayers.
AuthorsGraham I, Gagné J, Silvius JR
JournalBiochemistry
PubMed ID4084567
'The effects of calcium on the mixing of synthetic diacylphosphatidylcholines (PC''s) and diacylphosphatidylethanolamines (PE''s) with the corresponding phosphatidic acids (PA''s) have been examined by high-sensitivity differential scanning calorimetry and by measurements of the fluorescence of labeled PA or PC species in PA-PC bilayers. Calorimetrically derived phase diagrams for dimyristoyl- and ... More
Resonance energy transfer assay of protein-mediated lipid transfer between vesicles.
AuthorsNichols JW, Pagano RE
JournalJ Biol Chem
PubMed ID6853520
Comparative lateral diffusion of fluorescent lipid analogues in phospholipid multibilayers.
AuthorsDerzko Z, Jacobson K
JournalBiochemistry
PubMed ID7470449
Epifluorescence microscopic observation of monolayers of dipalmitoylphosphatidylcholine: dependence of domain size on compression rates.
AuthorsNag K, Boland C, Rich N, Keough KM
JournalBiochim Biophys Acta
PubMed ID1911829
A fluorescence microscopic technique was used to observe phase transitions in monolayers of DPPC. The sizes of the domain structures observed were found to be dependent on the rate of compression of the monolayer. The distribution of domain sizes for different rates of compression were unimodal, but the scatter in ... 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
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
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
A fluorescence-based high performance liquid chromatographic method for the characterization of palmitoyl acyl transferase activity.
AuthorsVarner AS, De Vos ML, Creaser SP, Peterson BR, Smith CD
JournalAnal Biochem
PubMed ID12234477
Although protein palmitoylation is essential for targeting many important signaling proteins to the plasma membrane, the mechanism by which palmitoylation occurs is uncharacterized, since the enzyme(s) responsible for this modification remain unidentified. To study palmitoyl acyl transferase (PAT) activity, we developed an in vitro palmitoylation (IVP) assay using a fluorescently ... More
Confocal imaging of time-dependent internalization and localization of NBD-PC in intact rat lungs.
AuthorsChinoy MR, Fisher AB, Shuman H
JournalAm J Physiol
PubMed ID8023961
Morphological localization of a fluorescent analogue of phosphatidylcholine (PC), C12-NBD-PC ([1-palmitoyl-2-[12-(7-nitro-2-1,3-benzoxadiazol-4-yl) amino] dodecanoyl]PC) was evaluated in adult rat lung. Fluorescence was observed at various times after intratracheal liposome instillation. Fluorometric analysis of C12-NBD-PC in lung homogenate showed an initial rapid uptake followed by a more gradual uptake phase. Microscopic observation ... More
Differential partitioning of pulmonary surfactant protein SP-A into regions of monolayers of dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylcholine/dipalmitoylphosphatidylglycerol.
AuthorsRuano ML, Nag K, Worthman LA, Casals C, Pérez-Gil J, Keough KM
JournalBiophys J
PubMed ID9512012
The interaction of the pulmonary surfactant protein SP-A fluorescently labeled with Texas Red (TR-SP-A) with monolayers of dipalmitoylphosphatidylcholine (DPPC) and DPPC/dipalmitoylphosphatidylglycerol 7:3 w/w has been investigated. The monolayers were spread on aqueous subphases containing TR-SP-A. TR-SP-A interacted with the monolayers of DPPC to accumulate at the boundary regions between liquid ... More
Fluorescently labeled pulmonary surfactant protein C in spread phospholipid monolayers.
AuthorsNag K, Perez-Gil J, Cruz A, Keough KM
JournalBiophys J
PubMed ID8804608
Pulmonary surfactant, a lipid-protein complex, secreted into the fluid lining of lungs prevents alveolar collapse at low lung volumes. Pulmonary surfactant protein C (SP-C), an acylated, hydrophobic, alpha-helical peptide, enhances the surface activity of pulmonary surfactant lipids. Fluorescein-labeled SP-C (F-SP-C) (3, 6, 12 wt%) in dipalmitoylphosphatidylcholine (DPPC), and DPPC:dipalmitoylphosphatidylglycerol (DPPG) ... More
Shape change and physical properties of giant phospholipid vesicles prepared in the presence of an AC electric field.
AuthorsMathivet L, Cribier S, Devaux PF
JournalBiophys J
PubMed ID8785271
Giant unilamellar vesicles with diameters ranging from 10 to 60 microns were obtained by the swelling of phospholipid bilayers in water in the presence of an AC electric field. This technique leads to a homogeneous population of perfectly spherical and unilamellar vesicles, as revealed by phase-contrast optical microscopy and freeze-fracture ... More
Transport of exogenous fluorescent phosphatidylserine analogue to the Golgi apparatus in cultured fibroblasts.
AuthorsKobayashi T, Arakawa Y
JournalJ Cell Biol
PubMed ID2010461
We have examined intracellular transport and metabolism of the fluorescent analogue of phosphatidylserine, 1-palmitoyl-2-(N-[12[(7-nitrobenz-2-oxa-1,3-diazole-4-yl)amino] dodecanoyl])-phosphatidylserine ([palmitoyl-C12-NBD]-PS) in cultured fibroblasts. When monolayer cultures were incubated with liposomes containing (palmitoyl-C12-NBD)-PS at 37 degrees C, fluorescent PS was transported to the Golgi apparatus. NBD-containing analogues of phosphatidylcholine, phosphatidylethanolamine (PE), or phosphatidic acid did ... More
Functional incorporation of integrins into solid supported membranes on ultrathin films of cellulose: impact on adhesion.
AuthorsGoennenwein S, Tanaka M, Hu B, Moroder L, Sackmann E
JournalBiophys J
PubMed ID12829518
Biomimetic models of cell surfaces were designed to study the physical basis of cell adhesion. Vesicles bearing reconstituted blood platelet integrin receptors alpha(IIb)beta(3) were spread on ultrathin films of cellulose, forming continuous supported membranes. One fraction of the integrin receptors, which were facing their extracellular domain toward the aqueous phase, ... More
Role of clathrin- and actin-dependent endocytotic pathways in lung phospholipid uptake.
AuthorsRückert P, Bates SR, Fisher AB
JournalAm J Physiol Lung Cell Mol Physiol
PubMed ID12611816
We evaluated the contribution of endocytotic pathways to pulmonary uptake of surfactant lipids from the alveolar space. Resting and stimulated 8-bromoadenosine 3',5'-cyclic monophosphate (8-Br-cAMP) uptake of unilamellar liposomes labeled with either [(3)H]dipalmitoylphosphatidylcholine ([(3)H]DPPC) or 1-palmitoyl-2-[12-(7-nitro-2-1,3-benzoxadiazol-4-yl) amino] dodecanoyl-phosphatidylcholine (NBD-PC) was studied in isolated perfused rat lungs and isolated type II cells. ... More
Dynamics of membrane penetration of the fluorescent 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD) group attached to an acyl chain of phosphatidylcholine.
AuthorsHuster D, Müller P, Arnold K, Herrmann A
JournalBiophys J
PubMed ID11159449
Location and dynamic reorientation of the fluorophore 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD) covalently attached to a short (C6) or a long (C12) sn2 acyl chain of a phosphatidylcholine molecule was investigated by fluorescence and solid-state NMR spectroscopy. 2H NMR lipid chain order parameters indicate a perturbation of the phospholipid packing density in the ... More
Uptake and cellular localization of exogenous lipids by Giardia lamblia, a primitive eukaryote.
AuthorsStevens TL, Gibson GR, Adam R, Maier J, Allison-Ennis M, Das S
JournalExp Parasitol
PubMed ID9207743
Giardia lamblia trophozoites are unable to carry out de novo lipid synthesis. It is therefore likely that lipids are acquired from the small intestine of the host, in which the trophozoites are exposed to free and conjugated fatty acids, various sterols, phospholipids, bile acids, and bile-lipid mixed micelles. Here we ... More
Electrostatic properties of membranes containing acidic lipids and adsorbed basic peptides: theory and experiment.
AuthorsMurray D, Arbuzova A, Hangyás-Mihályné G, Gambhir A, Ben-Tal N, Honig B, McLaughlin S
JournalBiophys J
PubMed ID10585939
The interaction of heptalysine with vesicles formed from mixtures of the acidic lipid phosphatidylserine (PS) and the zwitterionic lipid phosphatidylcholine (PC) was examined experimentally and theoretically. Three types of experiments showed that smeared charge theories (e.g., Gouy-Chapman-Stern) underestimate the membrane association when the peptide concentration is high. First, the zeta ... More
Spectroscopic and ionization properties of N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-labeled lipids in model membranes.
AuthorsChattopadhyay A, London E
JournalBiochim Biophys Acta
PubMed ID3337814
The spectroscopic and ionization properties of various lipids labeled with the 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD) group have been studied in model membranes using fluorescence, absorbance and electrophoretic mobility measurements. Electrophoretic measurements show that the NBD group is uncharged at neutral pH. However, at high pH, hydroxyl addition or deprotonation occurs with a ... More
Release of phospholipids from erythrocyte membranes by taurocholate is determined by their transbilayer orientation and hydrophobic backbone.
AuthorsWüstner D, Pomorski T, Herrmann A, Müller P
JournalBiochemistry
PubMed ID9836604
Bile salts mediate a specific release of phosphatidylcholine (PC) from the canalicular membrane into the bile fluid. We utilized human red blood cells (RBC) as a model system to study the release of endogenous phospholipids as well as phospholipid analogues from plasma membranes in the presence of the bile salt ... More
A phosphatidylinositol 4-kinase pleckstrin homology domain that binds phosphatidylinositol 4-monophosphate.
AuthorsStevenson JM, Perera IY, Boss WF
JournalJ Biol Chem
PubMed ID9712908
Pleckstrin homology (PH) domains are found in many proteins involved in signal transduction, including the family of large molecular mass phosphatidylinositol (PI) 4-kinases. Although the exact function of these newly discovered domains is unknown, it is recognized that they may influence enzyme regulation by binding different ligands. In this study, ... More
Differential determination of phospholipase A(2) and PAF-acetylhydrolase in biological fluids using fluorescent substrates.
AuthorsKitsiouli EI, Nakos G, Lekka ME
JournalJ Lipid Res
PubMed ID10588961
The purpose of the present study was the development and evaluation of a fluorimetric method for the screening and differential determination of phospholipase A(2) and PAF-acetylhydrolase in bronchoalveolar lavage (BAL) fluid and serum. Phospholipase A(2) was determined using C(12)-NBD-PC in the presence of Ca(2+), from the slope of the fluorescence ... More
Extension of the parallax analysis of membrane penetration depth to the polar region of model membranes: use of fluorescence quenching by a spin-label attached to the phospholipid polar headgroup.
AuthorsAbrams FS, London E
JournalBiochemistry
PubMed ID8399232
The parallax method is a method by which the depth of fluorescent molecules within a membrane is calculated from the ratio of quenching induced by two spin-labeled phospholipids at different depths. In this report, the method is extended to measurements of depth in the polar headgroup region of the membrane ... More
Asymmetric lateral mobility of phospholipids in the human erythrocyte membrane.
AuthorsMorrot G, Cribier S, Devaux PF, Geldwerth D, Davoust J, Bureau JF, Fellmann P, Herve P, Frilley B
JournalProc Natl Acad Sci U S A
PubMed ID3462734
The fluorescent phospholipid 1-acyl-2-[12-(7-nitrobenz-2-oxa-1,3-diazol-4- yl)aminododecanoyl]phosphatidylcholine (NBD-phosphatidylcholine) and the corresponding aminophospholipid derivatives (NBD-phosphatidylethanolamine and NBD-phosphatidylserine) were introduced in the human erythrocyte membrane by a nonspecific phospholipid exchange protein purified from corn. The lateral mobility of the fluorescent phospholipids was measured by using an extension of the classical photobleaching recovery technique that ... More
Control of a redox reaction on lipid bilayer surfaces by membrane dipole potential.
AuthorsAlakoskela JI, Kinnunen PK
JournalBiophys J
PubMed ID11159402
Nitro-2,1,3-benzoxadiazol-4-yl (NBD) group is a widely used, environment-sensitive fluorescent probe. The negatively charged dithionite rapidly reduces the accessible NBD-labeled lipids in liposomes to their corresponding nonfluorescent derivatives. In this study both the phospholipid headgroup and acyl chain NBD-labeled L-alpha-1,2-dipalmitoyl-sn-glycero-3-phospho-[N-(4-nitrobenz-2-oxa-1,3-diazole)-ethanolamine] (DPPN) and 1-acyl-2-[12-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]dodecanoyl]-sn-glycero-3-phosphocholine (NBD-PC), respectively, were employed. The correlation of both ... More
Adipocyte fatty acid-binding protein: interaction with phospholipid membranes and thermal stability studied by FTIR spectroscopy.
AuthorsGericke A, Smith ER, Moore DJ, Mendelsohn R, Storch J
JournalBiochemistry
PubMed ID9204877
Fatty acid-binding proteins (FABPs) found in many tissues constitute a family of low molecular weight proteins that are suggested to function as intracellular transporters of fatty acids. Studies of the transfer kinetics of fluorescent anthroyloxy-labeled long-chain fatty acids from FABP to model membranes led to the suggestion that the FABPs, ... More
Fluorescence measurements of fusion between human erythrocytes induced by poly(ethylene glycol).
AuthorsHuang SK, Hui SW
JournalBiophys J
PubMed ID2291937
The kinetics of poly(ethylene glycol) (PEG)-induced fusion between intact human erythrocytes was continuously monitored by a fluorescence lipid mixing method, utilizing the dequenching of the fluorescence probe, 1-oleoyl-2-[12-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]dodecanoyl ] phosphatidylcholine (C12-NBD-PC). The steady-state fluorescence intensity was detected from the surface of cells in a monolayer on an alcian blue-coated glass ... More
Annexin A4 reduces water and proton permeability of model membranes but does not alter aquaporin 2-mediated water transport in isolated endosomes.
AuthorsHill WG, Kaetzel MA, Kishore BK, Dedman JR, Zeidel ML
JournalJ Gen Physiol
PubMed ID12695484
Annexin A4 (Anx4) belongs to a ubiquitous family of Ca2+-dependent membrane-binding proteins thought to be involved in membrane trafficking and membrane organization within cells. Anx4 localizes to the apical region in epithelia; however, its physiological role is unclear. We show that Anx4 exhibited binding to liposomes (phosphatidylcholine:phosphatidylserine, 1:1) in the ... More
Differential effects of surfactant protein A on regional organization of phospholipid monolayers containing surfactant protein B or C.
AuthorsTaneva SG, Keough KM
JournalBiophys J
PubMed ID11023905
Epifluorescence microscopy combined with a surface balance was used to study monolayers of dipalmitoylphosphatidylcholine (DPPC)/egg phosphatidylglycerol (PG) (8:2, mol/mol) plus 17 wt % SP-B or SP-C spread on subphases containing SP-A in the presence or absence of 5 mM Ca(2+). Independently of the presence of Ca(2+) in the subphase, SP-A ... More
Adsorption of pulmonary surfactant protein SP-A to monolayers of phospholipids containing hydrophobic surfactant protein SP-B or SP-C: potential differential role for tertiary interaction of lipids, hydrophobic proteins, and SP-A.
AuthorsTaneva SG, Keough KM
JournalBiochemistry
PubMed ID10821681
Surface balance techniques were used to study the interactions of surfactant protein SP-A with monolayers of surfactant components preformed at the air-water interface. SP-A adsorption into the monolayers was followed by monitoring the increase in the surface pressure Deltapi after injection of SP-A beneath the films. Monolayers of dipalmitoylphosphatidylcholine (DPPC):egg ... More
Visualization of lateral phases in cholesterol and phosphatidylcholine monolayers at the air/water interface--a comparative study with two different reporter molecules.
AuthorsSlotte JP, Mattjus P
JournalBiochim Biophys Acta
PubMed ID7811742
This study has compared two chemically distinct NBD-lipids with regard to their partitioning properties into lateral phases of pure and mixed cholesterol/phosphatidylcholine monolayers. Pure NBD-cholesterol (22-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-23,24-bisnor++ +-5-cholen-3-ol), which has the NBD-function in the sterol side chain (at carbon 22), gave a liquid-expanded force-area isotherm on water at 22 degrees C ... More
Phase transitions in films of lung surfactant at the air-water interface.
AuthorsNag K, Perez-Gil J, Ruano ML, Worthman LA, Stewart J, Casals C, Keough KM
JournalBiophys J
PubMed ID9635752
Pulmonary surfactant maintains a putative surface-active film at the air-alveolar fluid interface and prevents lung collapse at low volumes. Porcine lung surfactant extracts (LSE) were studied in spread and adsorbed films at 23 +/- 1 degrees C using epifluorescence microscopy combined with surface balance techniques. By incorporating small amounts of ... More
Influence of obstacles on lipid lateral diffusion: computer simulation of FRAP experiments and application to proteoliposomes and biomembranes.
AuthorsSchram V, Tocanne JF, Lopez A
JournalEur Biophys J
PubMed ID7530654
Fluorescence Recovery After Photobleaching experiments were simulated using a computer approach in which a membrane lipid leaflet was mimicked using a triangular lattice obstructed with randomly distributed immobile and non-overlapping circular obstacles. Influence of the radius r and area fraction c of these obstacles and of the radius R of ... More
Phosphatidylcholine and phosphatidylethanolamine behave as substrates of the human MDR1 P-glycoprotein.
AuthorsBosch I, Dunussi-Joannopoulos K, Wu RL, Furlong ST, Croop J
JournalBiochemistry
PubMed ID9153408
The multidrug resistant cell line CEM/VBL300 and the parental CEM T-lymphoblastic cell line from which it was derived were used to study the accumulation of fluorescent phospholipid analogs of phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylserine (PS). The fluorescent analogs NBD-PC, NBD-PE, and NBD-PS and [3H]PC were delivered in liposomes prepared ... More
Epifluorescence microscopic studies of monolayers containing mixtures of dioleoyl- and dipalmitoylphosphatidylcholines.
AuthorsNag K, Keough KM
JournalBiophys J
PubMed ID8241382
Monolayers of dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylcholine (DOPC), and some mixtures of these lipids were investigated using an epifluorescence microscopic surface balance. Monolayers were visualized at 23 +/- 1 degree C through the fluorescence of 1 mol% of two different fluorescent probes, 1-palmitoyl-2-(12-[(7-nitro-2-1,3-benzoxadizole-4- yl)amino]dodecanoyl)phosphatidylcholine (NBD-PC), which partitions into the liquid expanded (LE) ... More
Cholesterol efflux to high-density lipoproteins and apolipoprotein A-I phosphatidylcholine complexes is inhibited by ethanol: role of apolipoprotein structure and cooperative interaction of phosphatidylcholine and cholesterol.
AuthorsAvdulov NA, Chochina SV, Igbavboa U, Wood WG
JournalBiochemistry
PubMed ID10956052
There is a substantial body of evidence showing that moderate alcohol consumption is associated with a reduced risk of cardiovascular morbidity and mortality. One of the factors thought to contribute to this reduction in risk is an increase in the level of high-density lipoproteins (HDL) correlated with alcohol consumption. However, ... 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
Fluorescence assay for phospholipid membrane asymmetry.
AuthorsMcIntyre JC, Sleight RG
JournalBiochemistry
PubMed ID1751498
Highly fluorescent 7-nitro-2,1,3-benzoxadiazol-4-yl-lipid (NBD-lipid) analogues are widely used to examine lipid transport and membrane structure. We have developed a method for chemically modifying NBD-labeled lipids in both artificial and biological membranes. This was achieved by treating fluorescently labeled membranes with dithionite (S2O4(-2)). When small unilamellar vesicles containing NBD-labeled phospholipids were ... More
Pore-forming peptides induce rapid phospholipid flip-flop in membranes.
AuthorsFattal E, Nir S, Parente RA, Szoka FC
JournalBiochemistry
PubMed ID8204607
A kinetic model for pore-mediated and perturbation-mediated flip-flop is presented and used to characterize the mechanism of peptide-induced phospholipid flip-flop in bilayers. The model assumes that certain peptides can bind to and aggregate within the membrane. When the aggregate attains a critical size (M peptides), a channel is created that ... More
Multiple mechanisms linked to platelet activation result in lysophosphatidic acid and sphingosine 1-phosphate generation in blood.
AuthorsSano T, Baker D, Virag T, Wada A, Yatomi Y, Kobayashi T, Igarashi Y, Tigyi G
JournalJ Biol Chem
PubMed ID11929870
Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (Sph1P) production was examined in vitro under conditions that simulated blood clotting. Several approaches were utilized to elucidate the metabolic pathways. 1) Platelet phospholipids were labeled using [32P]orthophosphate, and the production of [32P]Sph1P and LPA was examined. Thrombin stimulation of platelets resulted in rapid ... More
Cholesterol in condensed and fluid phosphatidylcholine monolayers studied by epifluorescence microscopy.
AuthorsWorthman LA, Nag K, Davis PJ, Keough KM
JournalBiophys J
PubMed ID9168032
Epifluorescence microscopy was used to investigate the effect of cholesterol on monolayers of dipalmitoylphosphatidylcholine (DPPC) and 1 -palmitoyl-2-oleoyl phosphatidylcholine (POPC) at 21 +/- 2 degrees C using 1 mol% 1-palmitoyl-2-[12-[(7-nitro-2-1, 3-benzoxadizole-4-yl)amino]dodecanoyl]phosphatidylcholine (NBD-PC) as a fluorophore. Up to 30 mol% cholesterol in DPPC monolayers decreased the amounts of probe-excluded liquid-condensed (LC) ... More
A 29-kDa protein associated with p67phox expresses both peroxiredoxin and phospholipase A2 activity and enhances superoxide anion production by a cell-free system of NADPH oxidase activity.
AuthorsLeavey PJ, Gonzalez-Aller C, Thurman G, Kleinberg M, Rinckel L, Ambruso DW, Freeman S, Kuypers FA, Ambruso DR
JournalJ Biol Chem
PubMed ID12121978
Production of toxic oxygen metabolites provides a mechanism for microbicidal activity of the neutrophil. The NADPH oxidase enzyme system initiates the production of oxygen metabolites by reducing oxygen to form superoxide anion (O(2)()). With stimulation of the respiratory burst, cytosolic oxidase components, p47(phox), p67(phox), and Rac, translocate to the phagolysomal ... More
Role of the helical domain in fatty acid transfer from adipocyte and heart fatty acid-binding proteins to membranes: analysis of chimeric proteins.
AuthorsLiou HL, Kahn PC, Storch J
JournalJ Biol Chem
PubMed ID11675390
The adipocyte and heart fatty acid-binding proteins (A- and HFABP) are members of a lipid-binding protein family with a beta-barrel body capped by a small helix-turn-helix motif. Both proteins are hypothesized to transport fatty acid (FA) to phospholipid membranes through a collisional process. Previously, we suggested that the helical domain ... More
Pulmonary surfactant protein A interacts with gel-like regions in monolayers of pulmonary surfactant lipid extract.
AuthorsWorthman LA, Nag K, Rich N, Ruano ML, Casals C, Pérez-Gil J, Keough KM
JournalBiophys J
PubMed ID11053138
Epifluorescence microscopy was used to investigate the interaction of pulmonary surfactant protein A (SP-A) with spread monolayers of porcine surfactant lipid extract (PSLE) containing 1 mol % fluorescent probe (NBD-PC) spread on a saline subphase (145 mM NaCl, 5 mM Tris-HCl, pH 6.9) containing 0, 0.13, or 0.16 microg/ml SP-A ... More
Extensive segregation of acidic phospholipids in membranes induced by protein kinase C and related proteins.
AuthorsBazzi MD, Nelsestuen GL
JournalBiochemistry
PubMed ID1868070
Protein kinase C and two other proteins with molecular masses of 64 and 32 kDa, purified from bovine brain, constitute a type of protein that binds a large number of calcium ions in a phospholipid-dependent manner. This study suggested that these proteins also induced extensive clustering of acidic phospholipids in ... More