Chemically induced phase separation in mixed vesicles containing phosphatidic acid. An optical study.
AuthorsGalla HJ, Sackmann E
JournalJ Am Chem Soc
PubMed ID1159213
Metabolism of 1-pyrenedecanoic acid and accumulation of neutral fluorescent lipids in cultured fibroblasts of multisystemic lipid storage myopathy.
AuthorsRadom J, Salvayre R, Maret A, Nègre A, Douste-Blazy L
JournalBiochim Biophys Acta
PubMed ID3607075
'The lipid metabolism in cultured fibroblasts from multisystemic (type 3) lipid storage myopathy and controls has been studied through pulse-chase experiments using 1-pyrenedecanoic acid as precursor. The uptake of 1-pyrenedecanoic acid was not significantly different in multisystemic lipid storage myopathy and control fibroblasts. The amount of fluorescent lipids synthesized by ... More
Lipid structural reorganization induced by the pancreatic lipase cofactor, procolipase.
AuthorsMomsen WE, Momsen MM, Brockman HL
JournalBiochemistry
PubMed ID7766639
'Pancreatic colipase and its precursor, procolipase, facilitate interfacial lipid hydrolysis catalyzed by pancreatic lipase. To better understand how procolipase functions, its interactions with mixed-lipid monolayers at the argon-buffer interface have been characterized. The lipid mixtures consisted of 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine and either 1,3-dioleoylglycerol, a model lipase substrate, or 13,16-cis,cis-docosadienoic acid, a model ... More
Changes in lipid distribution and dynamics in degranulated rat liver rough endoplasmic reticulum due to the membrane attachment of polyribosomes.
AuthorsKaihovaara P, Raulo E, Kinnunen PK
JournalBiochemistry
PubMed ID1883824
'Binding of rat liver polyribosomes to homologous degranulated rough endoplasmic reticulum (dRER) labeled with 10-(pyren-1-yl)decanoic acid (PDA) was studied. As a consequence of the membrane association of polysomes, the excimer/monomer fluorescence intensity ratios (Ie/Im) decreased, thus indicating alterations in the dynamics and organization of lipids. These fluorescence changes were complete ... More
Excimer-forming lipids in membrane research.
AuthorsGalla HJ, Hartmann W
JournalChem Phys Lipids
PubMed ID7418114
'Pyrenedecanoic acid and pyrene lecithin are optical probes well suited to investigate lipid bilayer membranes. The method is based on the determination of the formation of excited dimers or excimers. The rate of excimer formation yields information on the dynamic molecular properties of artificial as well as of natural membranes. ... More
C(60) and water-soluble fullerene derivatives as antioxidants against radical-initiated lipid peroxidation.
AuthorsWang IC, Tai LA, Lee DD, Kanakamma PP, Shen CK, Luh TY, Cheng CH, Hwang KC
JournalJ Med Chem
PubMed ID10579823
'C(60), vitamin E, and three C(60) derivatives (polar 1 and water-soluble C(3)/D(3)C(60)s) were examined for their antioxidant effects on prevention of lipid peroxidation induced by superoxide and hydroxyl radicals. The protection effect on lipid peroxidation was found to be in the sequence: C(60) >/= vitamin E > 1 > none, ... More
Association to HeLa cells and surface behavior of exogenous gangliosides studied with a fluorescent derivative of GM1.
AuthorsMasserini M, Giuliani A, Palestini P, Acquotti D, Pitto M, Chigorno V, Tettamanti G
JournalBiochemistry
PubMed ID2110827
'Cultured HeLa cells were incubated with pyrene-GM1/3H-radiolabeled GM1 ganglioside (1:4 M/M) mixtures for various times. The process of association of pyrene-GM1 with cells was qualitatively and quantitatively the same as that of 3H-GM1. The pyrene-GM1 and 3H-GM1 proportions in the various forms of association with cells were similar to that ... More
The lateral distribution of pyrene-labeled sphingomyelin and glucosylceramide in phosphatidylcholine bilayers.
AuthorsHresko RC, Sugár IP, Barenholz Y, Thompson TE
JournalBiophys J
PubMed ID3593870
'The lateral distribution of N-[10(1-pyrenyl)decanoyl]-sphingomyelin (PyrSPM) and N-[10(1-pyrenyl)decanoyl]-glucocerebroside (PyrGlcCer) was studied in multilamellar vesicles of 1,2-dipalmitoyl-, 1,2-dimyristoyl-, and 1-palmitoyl-2-oleoyl-phosphatidylcholine (DPPC, DMPC, and POPC, respectively) under anaerobic conditions by determining the excimer-to-monomer fluorescence intensity ratio (E/M) as a function of temperature. The E/M(T) curves for PyrSPM and PyrGlcCer in the three ... More
Interaction of the mitochondria-targeted antioxidant MitoQ with phospholipid bilayers and ubiquinone oxidoreductases.
'MitoQ(10) is a ubiquinone that accumulates within mitochondria driven by a conjugated lipophilic triphenylphosphonium cation (TPP(+)). Once there, MitoQ(10) is reduced to its active ubiquinol form, which has been used to prevent mitochondrial oxidative damage and to infer the involvement of reactive oxygen species in signaling pathways. Here we show ... More
Use of polymerized mixed liposomes to study interactions of phospholipase A2 with membranes.
AuthorsWu SK, Cho W
JournalBiochemistry
PubMed ID8268165
'Polymerized liposomes of thiol-based phospholipids, 1,2-bis[12-(lipoyloxy)dodecanoyl]-sn-glycero-3-phosphocholine (BLPC) and -phosphoglycerol (BLPG) were used to study interactions of several phospholipases A2 (PLA2) with membranes. Large liposomes (an average diameter of 100 +/- 10 nm) prepared from BLPC or BLPG were readily hydrolyzed by PLA2. Once polymerized, however, these liposomes were resistant to ... More
Hydrolysis of fluorescent pyrene-acyl esters by human pancreatic carboxylic ester hydrolase and bile salt-stimulated lipase.
AuthorsNegre-Salvayre A, Abouakil N, Lombardo D, Salvayre R
JournalLipids
PubMed ID2215086
'Fluorescent esters containing pyrenedecanoic acid (P10) or pyrenebutanoic (P4) acid (P4cholesterol, P10cholesterol, P4- and P10-containing triacylglycerols) were synthesized and used as substrates for human pancreatic carboxylic ester hydrolase and bile salt-stimulated lipase from human milk. Both enzymes were purified by immunoaffinity chromatography. All fluorescent pyrene derivatives were hydrolyzed by pancreatic ... More
A fluorometric determination of sphingomyelinase by use of fluorescent derivatives of sphingomyelin, and its application to diagnosis of Niemann-Pick disease.
AuthorsGatt S, Dinur T, Barenholz Y
JournalClin Chem
PubMed ID6243257
'We synthesized fluorescent derivatives of sphingomyelin (N-acyl-sphingosylphosphocholine) and used them as substrates for several sphingomyelinases. The following five fluorescent probes, each attached to the terminal carbon atom of the fatty acyl residue, were introduced into sphingomyelin: dansyl, pyrene, carbazole, 4-chloro-7-nitrobenz-2-oxa-1,3-diazole, and anthroic acid. We compared the rates at which the ... More
Thermodynamic and phase characterization of phosphatidylethanolamine and ganglioside GD1a mixtures.
AuthorsTsao YS, Freire E, Huang L
JournalBiochim Biophys Acta
PubMed ID3593711
'By employing diphenylhexatriene steady-state fluorescence anisotropy, pyrenedecanoic acid excimer formation, and high sensitivity scanning calorimetry we have demonstrated that the liposomes containing phosphatidylethanolamine (PE) and various mole fractions of ganglioside GD1a had a gel-liquid crystalline phase transition between 15 and 25 degrees C. Calorimetric measurements indicated that these phase transitions ... More
Effects of temperature and lipid composition on the serum albumin-induced aggregation and fusion of small unilamellar vesicles.
AuthorsSchenkman S, Araujo PS, Dijkman R, Quina FH, Chaimovich H
JournalBiochim Biophys Acta
PubMed ID7317422
'Small unilamellar vesicles of egg phosphatidylcholine (PC) or dimyristoylphosphatidylcholine, mixed with small unilamellar vesicles labelled with 2-(10-(1-pyrene)decanoyl)phosphatidylcholine, exhibit a constant average size and excimer to monomer (E/M) ratio for several hours when incubated at pH 3.6 at a temperature higher than the phase transition temperature (Tc) of the lipids. Addition ... More
Pyrene eximer mapping in cultured fibroblasts by ratio imaging and time-resolved microscopy.
AuthorsDix JA, Verkman AS
JournalBiochemistry
PubMed ID2331474
'The kinetics of pyrene eximer formation provide a measure of lateral diffusibility in bilayer membranes. Swiss 3T3 fibroblasts were labeled with pyrene, pyrenedecanoic acid (PDA) and 1,3-bis(1 pyrene) propane (BPP) by incubation in the presence of Pluronic F127. Single-cell emission spectra obtained by epifluorescence microscopy (excitation 350 nm) with photodiode ... More
Asymmetry of lipid dynamics in human erythrocyte membranes studied with permanent fluorophores.
AuthorsSchachter D, Cogan U, Abbott RE
JournalBiochemistry
PubMed ID7093236
'The fluorescence anisotropy and mean excited-state lifetime of 1,6-diphenyl-1,3,5-hexatriene, 12-(9-anthroyloxy)stearate, 2-(9-anthroyloxy)stearate, and pyrenedecanoic acid in the membranes of intact human erythrocytes, lysate suspensions, and ghost membranes were compared. The excited-state lifetime of each lipid fluorophore, estimated by single photon counting, is significantly shorter in the intact erythrocytes as compared to ... More
Uptake and intracellular degradation of fluorescent sphingomyelin by fibroblasts from normal individuals and a patient with Niemann-Pick disease.
AuthorsLevade T, Gatt S
JournalBiochim Biophys Acta
PubMed ID3567213
'Sphingomyelin, labelled with a fluorescent probe, pyrene, in the fatty acyl residue was associated with fetal calf serum; approx. 80% of the sphingomyelin was found in the low- and high-density lipoproteins. This was added to the growth medium of cultured human skin fibroblasts from normal individuals and a patient with ... More
Use of a fluorescent radiolabeled triacylglycerol as a substrate for lipoprotein lipase and hepatic triglyceride lipase.
AuthorsDousset N, Negre A, Salvayre R, Rogalle P, Dang QQ, Douste-Blazy L
JournalLipids
PubMed ID3172990
'A fluorescent radiolabeled triacylglycerol has been synthesized by using a fluorescent fatty acid (pyrene decanoic acid) and a radiolabeled oleic acid. This analog of the natural substrate, 1(3)pyrene decanoic-2,3 (1,2)-dioleoyl-sn-glycerol, has been tested as substrate for determining lipoprotein lipase and hepatic triacylglycerol lipase activities in post-heparin plasma. Optimal conditions for ... More
Membrane penetration of cytosolic phospholipase A2 is necessary for its interfacial catalysis and arachidonate specificity.
AuthorsLichtenbergova L, Yoon ET, Cho W
JournalBiochemistry
PubMed ID9760249
'To determine the mechanism of calcium-dependent membrane binding of cytosolic phospholipase A2 (cPLA2), we measured the interactions of cPLA2 with phospholipid monolayers and polymerizable mixed liposomes containing various phospholipids. In the presence of calcium, cPLA2 showed much higher penetrating power than secretory human pancreatic PLA2 toward anionic and electrically neutral ... More
Dietary triacylglycerol modulates sodium-dependent D-glucose transport, fluidity and fatty acid composition of rat small intestinal brush-border membrane.
AuthorsBrasitus TA, Dudeja PK, Bolt MJ, Sitrin MD, Baum C
JournalBiochim Biophys Acta
PubMed ID2923876
'Rats were maintained on nutritionally complete diets enriched in unsaturated (menhaden fish oil) or saturated (butter fat) triacylglycerols. After 4 weeks, the animals were killed, proximal small intestinal brush-border membranes were prepared, and examined and compared with respect to their lipid composition, molecular species of phosphatidylcholine, lipid fluidity and sodium-dependent ... More
Relative fluorescence of normal and acid lipase-deficient cultured fibroblasts following administration of pyrene decanoic acid.
AuthorsNègre A, Maret A, Douste-Blazy L, Gatt S, Salvayre R
JournalBiochim Biophys Acta
PubMed ID3382681
'Skin fibroblasts, derived from normal individuals or patients with Wolman''s disease (an autosomal recessive disorder due to acid lysosomal lipase deficiency) were incubated with the fluorescent fatty acid, pyrene-decanoic acid (P10). Measurements of the fluorescence intensities of the total lipid extracts indicated that equal quantities of P10 were incorporated into ... More
Membrane structural dynamics of plasma membranes of living human prostatic carcinoma cells differing in metastatic potential.
AuthorsCalderon RO, Grogan WM, Collins JM
JournalExp Cell Res
PubMed ID1893934
'Rigidity of the outer hemileaflet of the plasma membrane of two prostatic carcinoma cell lines with different metastatic potential, 1-LN and 1-LN-EMS-10, was assessed by steady-state anisotropy, using a battery of fluorescent probes. The "bulk" membrane rigidity sensed by diphenylhexatriene, trimethylammonio-DPH, 1-palmitoyl-2-[DPH-ethylcarbonyl]-phosphatidylcholine, and 10-pyrenedecanoic acid indicated slightly higher rigidity in ... More
Calorimetric and fluorescence characterization of interactions between cytochrome b5 and phosphatidylcholine bilayers.
AuthorsFreire E, Markello T, Rigell C, Holloway PW
JournalBiochemistry
PubMed ID6849876
'The interactions of cytochrome b5 with dimyristoylphosphatidylcholine and dipalmitoylphosphatidylcholine lipid bilayers have been studied with high-sensitivity differential scanning calorimetry and fluorescence spectroscopy. The incorporation of cytochrome b5 into large single lamellar vesicles causes a reduction in the enthalpy change associated with the lipid phase transition. Analysis of the dependence of ... More
Photosensitized killing of cultured fibroblasts from patients with peroxisomal disorders due to pyrene fatty acid-mediated ultraviolet damage.
AuthorsHoefler G, Paschke E, Hoefler S, Moser AB, Moser HW
JournalJ Clin Invest
PubMed ID1752949
'The influence of pyrene-fatty acids on the resistance of cells to ultraviolet (UV) radiation was investigated in cultured fibroblasts from patients with five types of peroxisomal disorders. All showed reduced survival compared to control. The effect varied with the biochemical defect involved and the chain length of the pyrene fatty ... More
Acyl-chain specificity and properties of cholesterol esterases from normal and Wolman lymphoid cell lines.
AuthorsNègre A, Salvayre R, Rogalle P, Dang QQ, Douste-Blazy L
JournalBiochim Biophys Acta
PubMed ID3030438
'Cholesteryl esters with various chain lengths of fatty acid, radioactive (C2-C18:1) and fluorescent (pyrene butanoic and decanoic acid, P4 and P10, respectively) were synthesized and their hydrolysis was investigated in lymphoid cell lines from normal subjects and from Wolman''s disease patients. The comparison of their hydrolysis showed that three cholesterol ... More
Erythrocyte, platelet and polymorphonuclear leukocyte membrane dynamic properties in essential hypertension.
AuthorsCaimi G
JournalClin Hemorheol Microcirc
PubMed ID9356784
'In subjects with essential hypertension we evaluated, respectively, the red cell membrane protein lateral mobility (obtained marking intact red blood cells with pyrene-3-maleimide (3-PM)), the erythrocyte membrane fluidity (obtained marking intact erythrocytes with 10-(1-pyrene) decanoic acid), the red cell membrane transverse fluidity gradient (obtained marking intact red blood cells with ... More
Fluorometric assay for pancreatic lipase.
AuthorsSalvayre R, Nègre A, Radom J, Douste-Blazy L
JournalClin Chem
PubMed ID3731446
'We report a new fluorometric assay in which a fluorescent triglyceride is used for determining lipase activity in serum, and we compare it with turbidimetric and radiometric methods. Because this fluorometric method is at least 50-fold more sensitive than the turbidimetric method, we have been able to develop a micromethod ... More
Joint determination by Brownian dynamics and fluorescence quenching of the in-depth location profile of biomolecules in membranes.
AuthorsFernandes MX, García de la Torre J, Castanho MA
JournalAnal Biochem
PubMed ID12137772
'The in-depth molar distribution function of fluorophores is revealed by a new methodology for fluorescence quenching data analysis in membranes. Brownian dynamics simulation was used to study the in-depth location profile of quenchers. A Lorentzian profile was reached. Since the Stern-Volmer equation is valid at every depth in the membrane ... More
Pyrenedecanoic acid and pyrene lecithin.
AuthorsGalla HJ, Hartmann W
JournalMethods Enzymol
PubMed ID7311843
Liposomal membranes. XIV. Fusion of liposomal membranes induced by polyisoprenoids as monitored by fluorescence quenching method.
AuthorsSunamoto J, Iwamoto K, Tezuka T, Kadosaki K, Kondo H
JournalChem Phys Lipids
PubMed ID6627530
Fusion of the single-walled liposomes of egg phosphatidylcholine as induced by the polyisoprenoids such as solanesol, trans-ethyl decaprenoate (EDP), coenzyme Q10, and dolichol has been investigated adopting the fluorescence quenching method. Relative efficiency of the polyisoprenoids employed on the induced fusion of liposomes was a sequence of solanesol less than ... More
Multimers of anionic amphiphiles mimic calmodulin stimulation of cyclic nucleotide phosphodiesterase.
AuthorsGietzen K, Xü YH, Galla HJ, Bader H
JournalBiochem J
PubMed ID6299276
Oleic acid, phosphatidylserine and pyrenedecanoic acid were found to activate calmodulin-deficient cyclic nucleotide phosphodiesterase at concentrations above their critical micellar concentration. In contrast with calmodulin these activators do not require the presence of Ca2+ for their action. It is shown that the size of phosphatidylserine vesicles is of crucial importance ... More
N-pyrene dodecanoyl sulfatide as membrane probe: a study of glycolipid dynamic behavior in model membranes.
AuthorsViani P, Galimberti C, Marchesini S, Cervato G, Cestaro B
JournalChem Phys Lipids
PubMed ID3342459
An N-linked pyrene-dodecanoyl sulfatide was employed to measure the ratio of excimer fluorescence to monomer fluorescence intensities (E/M). The E/M values provided information about both the dynamic behavior and the structural distribution of the labelled glycolipid in note dispersion of micellar sulfatides and multilamellar vesicles of different phospholipids. Most of ... More
Pyrene-labeled lipids: versatile probes of membrane dynamics in vitro and in living cells.
AuthorsPownall HJ, Smith LC
JournalChem Phys Lipids
PubMed ID2670287
Pyrene-labeled analogs of fatty acids have been studied as probes of lipid metabolism in vitro and in cultured cells. Procedures for the synthesis of complex pyrenyl lipids and the analytical methods for their separation and quantification are described. Pyrenyl-lipids have been used to quantify the relationship between lipid structure and ... More
Measurement of membrane fluidity of polymorphonuclear leukocytes by flow cytometry.
AuthorsMasuda M, Kuriki H, Komiyama Y, Nishikado H, Egawa H, Murata K
JournalJ Immunol Methods
PubMed ID3805741
A method was established for measuring membrane fluidity of polymorphonuclear leukocytes (PMN) by the excimer-forming lipid technique with pyrenedecanoic acid in flow cytometry. When cells were labeled, the use of 2-25 microM of pyrenedecanoic acid provided similar results. Neither the removal of the unincorporated pyrenedecanoic acid nor adjustment of PMN ... More
Import and fate of fluorescent analogs of oxidized phospholipids in vascular smooth muscle cells.
AuthorsMoumtzi A, Trenker M, Flicker K, Zenzmaier E, Saf R, Hermetter A,
JournalJ Lipid Res
PubMed ID17135656
Lipid oxidation is now thought to be an initiating and sustaining event in atherogenesis. Oxidatively fragmented phospholipids, namely 1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphocholine (PGPC) and 1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphocholine (POVPC), present in minimally modified LDL and atherosclerotic lesions, have been reported to elicit a wide range of pathophysiological responses in the cells of the vascular wall. Nevertheless, ... More
Enhancement of fluorescence of pyrene-containing lipids by polar media, detergents and phospholipids.
AuthorsLevade T, Salvayre R, Gatt S
JournalExperientia
PubMed ID3653339
The fluorescence intensities of a medium-chain fatty acid and of several amphiphilic lipids, each containing pyrene in covalent linkage, were enhanced considerably by: 1) Dissolving in mixtures of a polar solvent (e.g. methanol, ethanol, tetrahydrofuran or dimethylsulfoxide) and water; for each individual compound, a certain ratio of solvent to water ... More
Purification and spectroscopic properties of pyrene fatty acids.
AuthorsHresko RC, Markello TC, Barenholz Y, Thompson TE
JournalChem Phys Lipids
PubMed ID4085086
Pyrene fatty acids are routinely purified by silica based column chromatography and analyzed on thin-layer silica plates (H.-J. Galla et al., Chem. Phys. Lipids, 23 (1979) 239-251). Although pyrene decanoic acid runs as a single spot on thin-layer chromatography (TLC), gas-liquid chromatography (GC) of the methyl ester derivatives of a ... More
Pyrene-labeled lipids as tools in membrane biophysics and cell biology.
AuthorsSomerharju P
JournalChem Phys Lipids
PubMed ID12093535
Pyrene is one of the most frequently used lipid-linked fluorophores. Its most characteristic features are a long excited state lifetime and (local) concentration-dependent formation of excimers. Pyrene is also hydrophobic and thus does not significantly distort the conformation of the labeled lipid molecule. These characteristics make pyrene lipids well-suited for ... More
Free volume model for lipid lateral diffusion coefficients. Assessment of the temperature dependence in phosphatidylcholine and phosphatidylethanolamine bilayers.
AuthorsKing MD, Marsh D
JournalBiochim Biophys Acta
PubMed ID3768368
The lipid lateral diffusion coefficients, DT, in fluid-phase phosphatidylcholine and phosphatidylethanolamine bilayers have been analysed in terms of the free-volume diffusion model by fitting the expression: DT = AT exp[- B/(T - T0)] to the observed temperature dependence, where A, B and T0 are the parameters to be optimized. Application ... More
A sensitive and continuous fluorometric assay for phospholipase A2 using pyrene-labeled phospholipids in the presence of serum albumin.
AuthorsRadvanyi F, Jordan L, Russo-Marie F, Bon C
JournalAnal Biochem
PubMed ID2742139
Phospholipase A2 activity can be determined fluorometrically in the presence of serum albumin using phospholipids labeled at the sn-2-acyl position with 10-pyrenyldecanoic acid. In the water reaction medium 10-pyrene phospholipids form vesicles and the monomer fluorescence of the pyrene is negligible due to pyrene-pyrene interaction. Upon phospholipid hydrolysis 10-pyrenyldecanoic acids ... More
Long-chain-acyl-CoA synthetase and very-long-chain-acyl-CoA synthetase activities in peroxisomes and microsomes from rat liver. An enzymological study.
AuthorsLageweg W, Wanders RJ, Tager JM
JournalEur J Biochem
PubMed ID2007410
We have investigated the palmitic acid (C16:0) and cerotic acid (C26:0) activating activities in rat-liver microsomes and peroxisomes. The activation of the two fatty acids showed similar dependencies on ATP and coenzyme A, reflected in about equal apparent Km values both in microsomes and peroxisomes. In microsomes and peroxisomes similar ... More
A continuous fluorometric assay for phospholipase A(2) activity in brain cytosol.
AuthorsYarger DE, Patrick CB, Rapoport SI, Murphy EJ
JournalJ Neurosci Methods
PubMed ID11040375
Alterations in phospholipase A(2) (PLA(2)) activity have been implicated in Alzheimer disease and other neurological disorders, although brain PLA(2) activity is currently measured using lengthy, non-continuous assays. We describe herein a rapid, continuous assay in which we measured PLA(2) activity in mouse brain cytosol (CB-57). Brains were homogenized in HEPES ... More
Spontaneous and protein-mediated intermembrane transfer of the antiretroviral liponucleotide 3'-deoxythymidine diphosphate diglyceride.
Authorsvan Wijk GM, Gadella TW, Wirtz KW, Hostetler KY, van den Bosch H
JournalBiochemistry
PubMed ID1610834
Phospholipid conjugates of antiretroviral nucleosides show activity against the human immunodeficiency virus in vitro [Hostetler, K. Y., Stuhmiller, L. M., Lenting, H. B. M., Van den Bosch, H., & Richman, D. D. (1990) J. Biol. Chem. 265, 6112-6117]. In order to gain insight into the membrane association and the spontaneous ... More
Semisynthesis and purification of homogeneous plasmenylcholine molecular species.
AuthorsHan XL, Zupan LA, Hazen SL, Gross RW
JournalAnal Biochem
PubMed ID1317681
Methods for the efficient use of limiting amounts of fatty acid probes in the synthesis of individual molecular species of plasmenylcholine have been developed. Plasmenylcholine molecular species were synthesized through acylation of homogeneous 1-O-(Z)-hexadec-1'-enyl-sn-glycero-3-phosphocholine utilizing fatty acid anhydrides generated in situ from combined pools of reactant and recycled fatty acids ... More
High-performance liquid chromatography determination of bis(monoacylglycerol) phosphate and other lysophospholipids.
AuthorsLuquain C, Laugier C, Lagarde M, Pageaux JF
JournalAnal Biochem
PubMed ID11520030
Bis(monoacylglycerol) phosphate (BMP) is a very minor component of the phospholipid (PL) fraction in rat uterine stromal cell cultures (U(III) cells). Under several culture conditions, including the addition of (n-3) or (n-6) polyunsaturated fatty acids, BMP selectively accumulates docosahexaenoic acid (DHA). We have recently described the structure of this PL, ... More
Glucocerebroside transfer between phosphatidylcholine bilayers.
AuthorsCorrea-Freire MC, Barenholz Y, Thompson TE
JournalBiochemistry
PubMed ID7074080
We have studied the kinetics of transfer of glucocerebroside between phospholipid bilayers by using pyrene and 3H-labeled glucocerebroside incorporated into dimyristoylphosphatidylcholine (DMPC) and dipalmitoylphosphatidylcholine (DPPC) bilayers. Pyrene-labeled glucocerebroside (PyrCer) molecules are able to form an excited complex (eximer, E) between a PyrCer in the ground state and an excited monomer ... More
Metabolism of pyrenedecanoic acid in Epstein-Barr virus-transformed lymphoid cell lines from normal subjects and from a patient with multisystemic lipid storage myopathy.
AuthorsRadom J, Salvayre R, Negre A, Douste-Blazy L
JournalBiochim Biophys Acta
PubMed ID2550076
A lymphoid cell line has been established from a patient with multisystemic lipid storage myopathy and showed a major triacylglycerol storage, whereas the content of other neutral lipids and phospholipids was in the normal range. The metabolism of the triacylglycerols has been investigated in this lymphoid cell line from multisystemic ... More
Pyrene phospholipid as a biological fluorescent probe for studying fusion of virus membrane with liposomes.
AuthorsPal R, Barenholz Y, Wagner RR
JournalBiochemistry
PubMed ID2831956
We are using fluorescent endogenous phospholipids in virus membranes to study the factors that promote fusion on interaction with receptor membranes. To this end, vesicular stomatitis virus (VSV) grown in baby hamster kidney (BHK-21) cells was biologically labeled with fluorescent lipids, primarily phosphatidylcholine and phosphatidylethanolamine, derived from pyrene fatty acids. ... More
Transport of fluorescent derivatives of fatty acids into cultured human leukemic myeloid cells and their subsequent metabolic utilization.
AuthorsMorand O, Fibach E, Dagan A, Gatt S
JournalBiochim Biophys Acta
PubMed ID6954994
Transport of fluorescent derivatives of fatty acids across the cell membrane of cultured human leukemic myeloid cells (HL 60) and their subsequent metabolic utilization were studied. The rates of uptake of these derivatives and their incorporation into cellular lipids wer compared with that of radioactively labelled palmitic acid. Three groups ... More
Effect of hypothyroidism on the lipid composition and fluidity of rat colonic apical plasma membranes.
AuthorsBrasitus TA, Dudeja PK
JournalBiochim Biophys Acta
PubMed ID3128322
Prior studies have suggested that the lipid composition and lipid fluidity of cellular membranes of various organs are altered in the hypothyroid rat. To date, the effects of hypothyroidism on these parameters have not been examined in rat colonic apical plasma membranes. In the present experiments, male Sprague-Dawley rats were ... More
Fluorescent lipid probes in the study of viral membrane fusion.
AuthorsBlumenthal R, Gallo SA, Viard M, Raviv Y, Puri A
JournalChem Phys Lipids
PubMed ID12093534
Fluorescent lipid probes are widely used in the observation of viral membrane fusion, providing a sensitive method to study fusion mechanism(s). Due to the wealth of data concerning liposome fusion, a variety of fusion assays has been designed including fluorescent probe redistribution, fluorescence dequenching, fluorescence resonance energy transfer and photosensitized ... 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
Abnormal membrane fluidity as a cause of impaired functional dynamics of chemoattractant receptors on neonatal polymorphonuclear leukocytes: lack of modulation of the receptors by a membrane fluidizer.
AuthorsYasui K, Masuda M, Matsuoka T, Yamazaki M, Komiyama A, Akabane T, Hasui M, Kobayashi Y, Murata K
JournalPediatr Res
PubMed ID2845343
Membrane properties associated with chemoattractant-mediated cellular responsiveness of neonatal polymorphonuclear leukocytes (PMN) were analyzed using n-formylmethionyl-leucyl-phenylalanine. Inasmuch as aliphatic alcohols as a membrane fluidizer can enhance the chemoattractant binding and affect subsequent cellular responsiveness in adult PMN, neonatal PMN were studied for such properties by their treatment with iso-propyl alcohol, ... More
Evaluation of membrane physiology following fluorescence activated or magnetic cell separation.
AuthorsSeidl J, Knuechel R, Kunz-Schughart LA
JournalCytometry
PubMed ID10554157
BACKGROUND: Over the past decade, cell separation technology has become an important tool in various fields of cell biology allowing for the analysis or subsequent cultivation of specific cell subsets. The objective of the present study was to evaluate if the established sorting techniques fluorescence-activated (FACS) and magnetic cell separation ... More
Lateral organization of pyrene-labeled lipids in bilayers as determined from the deviation from equilibrium between pyrene monomers and excimers.
AuthorsBarenholz Y, Cohen T, Haas E, Ottolenghi M
JournalJ Biol Chem
PubMed ID8621705
In lipid bilayers, pyrene and pyrene-labeled lipids form excimers in a concentration-dependent manner. The aromatic amine N, N-diethylaniline (DEA), which has a high membrane-to-medium partition coefficient, quenches the monomers only, and therefore it is expected that under conditions in which the monomers are in equilibrium with the excimers due to ... More
Dissociation of cytochrome c from liposomes by histone H1. Comparison with basic peptides.
AuthorsRytömaa M, Kinnunen PK
JournalBiochemistry
PubMed ID8605203
The basic chromosomal protein histone H1 binds avidly to liposomes containing acidic phospholipids and with characteristics somewhat resembling the lipid association of cytochrome c (cyt c) [Koiv et al. (1995) Biochemistry 34, 8018-8027]. Membrane association of histone H1 strongly attenuates the lipid lateral diffusion in large unilamellar vesicles containing phosphatidylglycerol ... More
Metabolism of pyrenyl fatty acids in baby hamster kidney fibroblasts. Effect of the acyl chain length.
AuthorsKasurinen J, Somerharju P
JournalJ Biol Chem
PubMed ID1551867
Biosynthetic labeling of cellular lipids with a fluorescent pyrenyl fatty acid (PyrxFA) moiety was studied in order to assess the usefulness of this approach in the introduction of fluorescent lipid molecules to living cells for transport and metabolic studies. PyrxFAs containing 4-14 aliphatic carbons were added to the culture medium ... More
Hydrolysis of fluorescent pyrenetriacylglycerols by lipases from human stomach and gastric juice.
AuthorsNègre A, Salvayre R, Dousset N, Rogalle P, Dang QQ, Douste-Blazy L
JournalBiochim Biophys Acta
PubMed ID3196738
Fluorescent triacylglycerols containing pyrenedecanoic (P10) and pyrenebutanoic (P4) acids were synthesized and their hydrolysis by lipases from human gastric juice and stomach homogenate was investigated. The existence in stomach homogenate of four different lipolytic enzymes hydrolyzing fluorescent triacylglycerols is suggested by the comparison of various enzymatic properties: acyl chain length ... More
Spontaneous domain formation of phospholipase A2 at interfaces: fluorescence microscopy of the interaction of phospholipase A2 with mixed monolayers of lecithin, lysolecithin and fatty acid.
AuthorsReichert A, Ringsdorf H, Wagenknecht A
JournalBiochim Biophys Acta
PubMed ID1581331
Fluorescence microscopy has recently been proven to be an ideal tool to investigate the specific interaction of phospholipase A2 with oriented substrate monolayers. Using a dual labeling technique, it could be shown that phospholipase A2 can specifically attack and hydrolyze solid analogous L-alpha-DPPC domains. After a critical extent of monolayer ... More
Organization and dynamics of pyrene and pyrene lipids in intact lipid bilayers. Photo-induced charge transfer processes.
AuthorsBarenholz Y, Cohen T, Korenstein R, Ottolenghi M
JournalBiophys J
PubMed ID1883931
The dynamics of fluorescence quenching and the organization of a series of pyrene derivatives anchored in various depths in bilayers of phosphatidylcholine small unilamellar vesicles was studied and compared with their behavior in homogeneous solvent systems. The studies include characterization of the environmental polarity of the pyrene fluorophore based on ... More
Protease priming of neutrophil superoxide production. Effects on membrane lipid order and lateral mobility.
AuthorsKusner DJ, Aucott JN, Franceschi D, Sarasua MM, Spagnuolo PJ, King CH
JournalJ Biol Chem
PubMed ID1653237
Phagocyte superoxide (O2-) response is primed by a variety of physiologic compounds including the neutrophil secretory proteases cathepsin G and elastase. To study whether protease priming of neutrophil O2- response is related to changes in membrane physical state, we examined enzyme effects on the order and lateral mobility of lipid ... More
The phosphatidyl-myo-inositol anchor of the lipoarabinomannans from Mycobacterium bovis bacillus Calmette Guérin. Heterogeneity, structure, and role in the regulation of cytokine secretion.
AuthorsNigou J, Gilleron M, Cahuzac B, Bounéry JD, Herold M, Thurnher M, Puzo G
JournalJ Biol Chem
PubMed ID9287310
Lipoarabinomannans are major mycobacterial antigens capable of modulating the host immune response; however, the molecular basis underlying the diversity of their immunological properties remain an open question. In this study a new extraction and purification approach was successfully applied to isolate ManLAMs (lipoarabinomannans with mannosyl extensions) from bacillus Calmette Guérin ... More
Spectroscopic evidence for a preferential location of lidocaine inside phospholipid bilayers.
AuthorsFraceto LF, Pinto Lde M, Franzoni L, Braga AA, Spisni A, Schreier S, de Paula E
JournalBiophys Chem
PubMed ID12408938
We examined the effect of uncharged lidocaine on the structure and dynamics of egg phosphatidylcholine (EPC) membranes at pH 10.5 in order to assess the location of this local anesthetic in the bilayer. Changes in the organization of small unilamellar vesicles were monitored either by electron paramagnetic resonance (EPR)-in the ... More
Calcium-induced lipid phase separations and interactions of phosphatidylcholine/anionic phospholipid vesicles. Fluorescence studies using carbazole-labeled and brominated phospholipids.
AuthorsSilvius JR
JournalBiochemistry
PubMed ID2337575
A novel method that uses a carbazole-labeled fluorescent phosphatidylcholine, which partitions preferentially into liquid-crystalline lipid domains, to monitor the kinetics and the extents of thermotropic and ionotropic lateral phase separations in vesicles combining brominated and nonbrominated phosphatidylcholines (PCs), phosphatidic acids (PAs), and phosphatidylserines (PSs) is described. The calcium-induced segregation of ... More
Lateral and transversal diffusion and phase transitions in erythrocyte membranes. An excimer fluorescence study.
AuthorsGalla HJ, Luisetti J
JournalBiochim Biophys Acta
PubMed ID7353003
The lateral diffusion of the excimer-forming probe pyrene decanoic acid has been determined in erythrocyte membranes and in vesicles of the lipid extracts. The random walk of the probe molecules is characterized by their jump frequency, nu j, within the lipid matrix. At T = 35 degrees C a value ... More
Platelet agonists enhance the import of phosphatidylethanolamine into human platelets.
AuthorsEngelmann B, Schaipp B, Dobner P, Stoeckelhuber M, Kögl C, Siess W, Hermetter A
JournalJ Biol Chem
PubMed ID9774389
It is unknown whether the endocytosis-independent transfer of phospholipids from lipoproteins to platelets is regulated by platelet agonists such as thrombin. The movements of the choline phospholipids phosphatidylcholine and sphingomyelin (labeled with either 14C or the fluorescent pyrenedecanoic acid) between low density lipoproteins and platelets were unaffected by thrombin (0.5 ... More
1,2-Dimethylhydrazine-induced premalignant alterations in the S-adenosylmethionine/S-adenosylhomocysteine ratio and membrane lipid lateral diffusion of the rat distal colon.
AuthorsHalline AG, Dudeja PK, Brasitus TA
JournalBiochim Biophys Acta
PubMed ID3415996
Prior studies by our laboratory, utilizing the 1,2-dimethylhydrazine experimental model of colonic cancer, had shown that administration of this procarcinogen for 5 weeks was found to increase phospholipid methyltransferase activity and the fluidity of rat distal colonic brush-border membranes. The present studies were conducted to further explore these 'premalignant' colonic ... More
A continuous fluorometric assay for phospholipases using polymerized mixed liposomes.
AuthorsWu SK, Cho W
JournalAnal Biochem
PubMed ID7985786
A versatile continuous fluorometric assay for phospholipases A2, C, and D has been developed utilizing polymerized mixed liposomes made of pyrene-containing phospholipids (5 mol%) uniformly inserted in the polymerized liposomes of 1,2-bis[12-(lipoyloxy)dodecanoyl]-sn-glycero-3-phosphoglycerol (BLPG) and its derivatives. 1-Hexadecanoyl-2-(1-pyrenedecanoyl)-sn-glycero-3-phosphocholine was used for phospholipase A2 and N-(1-pyrenesulfonyl)-egg phosphatidyl ethanolamine for phospholipases C and ... More
Effect of docosahexaenoic acid on mouse mitochondrial membrane properties.
AuthorsStillwell W, Jenski LJ, Crump FT, Ehringer W
JournalLipids
PubMed ID9168456
Long-chain polyunsaturated (n-3) fatty acids have been proposed to be involved in a wide variety of biological activities. In this study, mitochondrial docosahexaenoic acid (DHA) levels were increased by either dietary manipulation or by fusing the mitochondria with phospholipid vesicles made from 1-stearoyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine (18:0/22:6 PC). The fused mitochondria exhibited a ... More
On two-dimensional passive random walk in lipid bilayers and fluid pathways in biomembranes.
AuthorsGalla HJ, Hartmann W, Theilen U, Sackmann E
JournalJ Membr Biol
PubMed ID40032
The lateral mobility of pyrene, pyrene decanoic acid, and 1-palmitoyl-2-pyrene decanoyl-phosphatidyl choline (pyrene lecithin) in lipid bilayers is determined by the excimer formation technique. This method is applied to vesicles of lecithins differing in chain length and in the degree of saturation of the hydrocarbon chains. These values are ... More