The peroxin pex3p initiates membrane assembly in peroxisome biogenesis.
AuthorsGhaedi K, Tamura S, Okumoto K, Matsuzono Y, Fujiki Y
JournalMol Biol Cell
PubMed ID10848631
'Rat cDNA encoding a 372-amino-acid peroxin was isolated, primarily by functional complementation screening, using a peroxisome-deficient Chinese hamster ovary cell mutant, ZPG208, of complementation group 17. The deduced primary sequence showed approximately 25% amino acid identity with the yeast Pex3p, thereby we termed this cDNA rat PEX3 (RnPEX3). Human and ... More
Disappearance of plasmalogens from membranes of animal cells subjected to photosensitized oxidation.
AuthorsMorand OH, Zoeller RA, Raetz CR
JournalJ Biol Chem
PubMed ID3403548
'Chinese hamster ovary cells (CHO-K1) photosensitized with 12-(1''-pyrene)dodecanoic acid (P12) are killed when exposed to long wavelength ultraviolet (UV) light (greater than 300 nm). Mutants deficient in plasmalogen biosynthesis are hypersensitive to this treatment. We now demonstrate that plasmenylethanolamine is rapidly and preferentially destroyed when CHO-K1 cells, photosensitized either with ... More
Small unilamellar vesicles are able to fuse with Mycoplasma capricolum cells.
AuthorsSalman M, Tarshis M, Rottem S
JournalBiochim Biophys Acta
PubMed ID1901498
'We have investigated the fusion characteristics of intact Mycoplasma capricolum cells and small unilamellar vesicles (SUV). The rate and extent of fusion was monitored continuously by octadecylrhodamine B (R18) fluorescence dequenching assay, as well as by intracellular contents mixing, and by sucrose density gradient analysis. The fusion of SUV with ... More
Fluorescence studies of the location and membrane accessibility of the palmitoylation sites of rhodopsin.
AuthorsMoench SJ, Moreland J, Stewart DH, Dewey TG
JournalBiochemistry
PubMed ID8180207
'Fluorescent fatty acid labels have been incorporated into the palmitoylation sites of rhodopsin and used to probe the membrane accessibility and location of these sites. The fluorescence properties of anthroyloxy and pyrenyl fatty acids bound to rhodopsin were investigated in a reconstituted vesicle system. Collisional quenching of fluorescence by stearic ... More
Interaction of partially structured states of acidic fibroblast growth factor with phospholipid membranes.
AuthorsMach H, Middaugh CR
JournalBiochemistry
PubMed ID7543282
'Although acidic fibroblast growth factor (aFGF) lacks a conventional signal sequence, it is often found complexed to sulfated proteoglycans on the external surface of cells. The protein also forms a "molten globule"-like state at neutral pH and physiological temperatures as well as at acidic pH in the presence of physiological ... More
Spontaneous and plasma factor-mediated transfer of pyrenyl cerebrosides between model and native lipoproteins.
AuthorsVia DP, Massey JB, Vignale S, Kundu SK, Marcus DM, Pownall HJ, Gotto AM
JournalBiochim Biophys Acta
PubMed ID3931685
'A series of pyrenyl glucocerebrosides was synthesized by reacylation of psychosine with pyrene-labeled fatty acids having 3-11 methylene units. When incorporated into model high-density lipoproteins consisting of dimyristoylphosphatidylcholine-apolipoprotein A-II complexes and incubated with unlabeled complexes, these lipids exhibited spontaneous transfer. Half times of transfer varied from 1.5 min to 365 ... 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
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
Fluorometry of turbid and absorbant samples and the membrane fluidity of intact erythrocytes.
AuthorsEisinger J, Flores J
JournalBiophys J
PubMed ID4016211
'In employing intrinsic or extrinsic fluorophores in the study of whole cells, or other strongly absorbant and/or scattering samples, the measured fluorescence intensity and polarization is seriously affected by absorption and scattering within the sample cuvet. These artifacts are analyzed and simple protocols are provided for overcoming them. An expression ... More
Correlation of the dispersion state of pyrene cerebroside sulfate and its uptake and degradation by cultured cells.
AuthorsViani P, Marchesini S, Cestaro B, Gatt S
JournalBiochim Biophys Acta
PubMed ID2923862
'This study aimed at increasing the efficiency and shortening the time required for administering cerebroside sulfate to cultured cells. For this purpose several modes of dispersion of a fluorescent derivative of cerebroside sulfate (sulfatide), in which the natural fatty acid has been replaced by pyrenedodecanoic acid (P12), were incubated with ... More
Diagnosis of arylsulfatase A deficiency in intact cultured cells using a fluorescent derivative of cerebroside sulfate.
AuthorsBach G, Dagan A, Herz B, Gatt S
JournalClin Genet
PubMed ID2885115
'A fluorescent derivative of cerebroside sulfate (12-(1-pyrene)dodecanoyl-sphingosylgalactosyl-0-3-sulfate (P12-sulfatide) has been synthesized as a potential substrate for the determination of cerebroside sulfatidase (or arylsulfatase A) activity. It was administered into cultured human skin fibroblasts and thereby utilized for the diagnosis of arylsulfatase A deficiency. Cultured skin fibroblasts from normal individuals and ... More
Pyrene-labeled gangliosides: micelle formation in aqueous solution, lateral diffusion, and thermotropic behavior in phosphatidylcholine bilayers.
AuthorsOllmann M, Schwarzmann G, Sandhoff K, Galla HJ
JournalBiochemistry
PubMed ID3676298
'By use of the excimer technique, the formation in aqueous solution of pyrene-labeled ganglioside micelles and their lateral diffusion and distribution in phosphatidylcholine membranes were investigated. For these studies 12-(1-pyrenyl)dodecanoic acid was covalently attached to the ceramide part of lysogangliosides GM1, GM2, GM3, GD1a, and GD1b. The 12-(1-pyrenyl)dodecanoic acid substitute ... More
Pyrenedodecanoylcarnitine and pyrenedodecanoyl coenzyme A: kinetics and thermodynamics of their intermembrane transfer.
'The intermembrane transfer kinetics and transition-state thermodynamics of pyrenedodecanoylcarnitine (PDC), pyrenedodecanoyl coenzyme A (PDCoA), and pyrenedodecanoic acid (PDA) were measured by observing the time-dependent decay in pyrene excimer fluorescence. Probe molecules transferred more slowly with an increase in vesicle size. Rates of PDC and PDA transfer were increased from a ... 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
Continuous spectrofluorometric measurements of uptake by cultured cells of 12-(1-pyrene)-dodecanoic acid from its complex with albumin.
AuthorsGatt S, Nahas N, Fibach E
JournalBiochem J
PubMed ID3178719
'Aqueous dispersions of 12-(1-pyrene)-dodecanoic acid (P12), a medium-chain fatty acid to which the fluorescent probe pyrene has been covalently linked, shows a considerable increase in fluorescence when the probe is introduced into a hydrophobic environment. This enables the uptake of P12 by liposomes and cells to be followed directly in ... More
Definition of the specific roles of lysolecithin and palmitic acid in altering the susceptibility of dipalmitoylphosphatidylcholine bilayers to phospholipase A2.
AuthorsHenshaw JB, Olsen CA, Farnbach AR, Nielson KH, Bell JD
JournalBiochemistry
PubMed ID9692961
'Bilayers composed of phosphatidylcholine initially resist catalysis by phospholipase A2. However, after a latency period, they become susceptible when sufficient reaction products (lysolecithin and fatty acid) accumulate in the membrane. Temperature near the main bilayer phase transition and calcium concentration modulate the effectiveness of the reaction products. The purpose of ... More
Flow cytofluorometric analysis of the uptake of the fluorescent fatty acid pyrene-dodecanoic acid by human peripheral blood cells.
AuthorsFibach E, Giloh H, Rachmilewitz EA, Gatt S
JournalCytometry
PubMed ID3208618
'The fluorescence activated cell sorter (FACS) was used for measuring the uptake of the fluorescent fatty acid derivative 12-(1-pyrene) dodecanoic acid (P12) by human peripheral blood cells. The results indicate that blood cells differ widely in their ability to take up P12, with polymorphonuclear cells showing the greatest uptake, followed ... More
Consumption of carotenoids in photosensitized oxidation of human plasma and plasma low-density lipoprotein.
AuthorsOjima F, Sakamoto H, Ishiguro Y, Terao J
JournalFree Radic Biol Med
PubMed ID8225019
'Human plasma and plasma low-density lipoprotein (LDL) were exposed to photoirradiation in the presence of methylene blue (water-soluble photosensitizer) or 12-(1-pyrene)dodecanoic acid (P-12, lipid-soluble photosensitizer). In methylene-blue-sensitized photooxidation of human plasma and LDL, endogenous carotenoids and tocopherols were consumed with the accumulation of cholesteryl ester hydroperoxides (CE-OOH). Xanthophylls (zeaxanthin and ... More
The lateral fluidity of erythrocyte membranes. Temperature and pressure dependence.
AuthorsEisinger J, Scarlata SF
JournalBiophys Chem
PubMed ID3440128
'The pressure and temperature dependence of the lateral and rotational fluidity of erythrocyte membranes was investigated by inserting the excimeric membrane probe 1''-pyrenedodecanoic acid (PDA) into the membranes of intact cells and measuring the probe excimer formation rate and the steady-state polarization of the monomer at pressures up to 2000 ... More
Uptake of fluorescent fatty acids by erythroleukemia cells. Effect of differentiation.
AuthorsFibach E, Nahas N, Giloh H, Gatt S
JournalExp Cell Res
PubMed ID3462004
'The uptake of a fluorescent derivative of a fatty acid (FDFA), 12-(1-pyrene) dodecanoic acid (P12) by murine erythroleukemia (MEL) cells was studied. Because of the intense fluorescence of the pyrene ring, the association of P12 with intact cells could be analysed using a fluorescence microscope or a fluorescence-activated cell sorter ... 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
Lysogangliosides: synthesis and use in preparing labeled gangliosides.
AuthorsSchwarzmann G, Sandhoff K
JournalMethods Enzymol
PubMed ID3600331
Pyrene dodecanoic acid coenzyme A ester: peroxisomal oxidation and chain shortening.
AuthorsGatt S, Bremer J, Osmundsen H
JournalBiochim Biophys Acta
PubMed ID3334862
Pyrenedodecanoyl-CoA was beta-oxidized by isolated rat liver peroxisomes at a rate which was about 50% of that observed with palmitoyl-CoA. Measurement of the quantity of NADH formed from a limiting amount of pyrenedodecanoyl-CoA suggested that it was subjected to two to three cycles of beta-oxidation. Pyrenedodecanoyl-CoA was a very poor ... 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
The conformation of pyrethroids bound to lipid bilayers.
AuthorsMichelangeli F, Robson MJ, East JM, Lee AG
JournalBiochim Biophys Acta
PubMed ID2207119
A number of pyrethroids have been synthesised containing bromine or iodine atoms in the acid and alcohol moieties. Quenching of the fluorescence of a pyrene-containing fatty acid in phospholipid bilayers has been used to measure the partitioning of the pyrethroids into the bilayers. Comparison of the intensity of the fluorescence ... 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
Heterogeneity of microsomal membrane fluidity: evaluation using intrinsic tryptophan energy transfer to pyrene probes.
AuthorsEngelke M, Behmann T, Ojeda F, Diehl HA
JournalChem Phys Lipids
PubMed ID7923478
Membrane fluidity measurements based on excimer formation of pyrene and pyrene derivatives as a measure of lateral diffusion yield a decreased fluidity in the presence of proteins [1-3]. It was the aim of our study to investigate whether the reduced excimer formation is due to a rigidifying effect of proteins ... 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
Induction of lipid storage in cultured leukemic myeloid cells by pyrene-dodecanoic acid.
AuthorsMorand O, Fibach E, Livni N, Gatt S
JournalBiochim Biophys Acta
PubMed ID6584182
When incubated for 1-3 days in the presence of the fatty acid analog, 12-(1-pyrene)dodecanoic acid, the neutral lipid content of cultured human leukemic myeloid cells increased considerably, while that of the phospholipids increased to a much lesser extent. Among the neutral lipids, di- and monoacylglycerols predominated and a considerable portion ... More
Fusion and lipid exchange in vesicles containing lipophilic spin labels.
AuthorsDefrise-Quertain F, Chatelain P, Ruysschaert JM, Delmelle M
JournalBiochim Biophys Acta
PubMed ID7093268
Lipophilic non-electrolyte spin labels greatly accelerate the fusion of unilamellar vesicles of dipalmitoylphosphatidylcholine when the system is maintained below the lipid phase transition. Differential scanning calorimetry and centrifugation measurements show that the transformed vesicles are large and probably unilamellar. Differential scanning calorimetry and fluorescence depolarization measurements were also carried out ... More
Regulation of sterol carrier protein gene expression by the forkhead transcription factor FOXO3a.
The SCP gene encodes two proteins, sterol carrier protein X (SCPx) and SCP2, that are independently regulated by separate promoters. SCPx has been shown to be the thiolase involved in the breakdown of branched-chain fatty acids and in the biosynthesis of bile acids. The in vivo function of SCP2 however ... More
Selective photosensitization of human leukemic cells by a pyrene-containing fatty acid.
AuthorsFibach E, Gatt S, Rachmilewitz EA
JournalExp Hematol
PubMed ID2303106
Irradiation with long-wave UV light (LUV) at 366 nm of cells that had been incubated with 12-(1-pyrene)dodecanoic acid (P12), a fatty acid derivative with a covalently linked pyrene nucleus, resulted in cytotoxicity. Using the in vitro established human cell lines HL-60 and U-937, we demonstrated that these leukemic cells are ... More
A high-performance liquid chromatographic method for the separation and quantification of acylated lipids containing pyrene fatty acids is described. The method is adapted from a procedure originally developed for the analysis of tissue lipids (Christie, W. W. (1985) J. Lipid Res. 26, 507-512). Pyrenyl lipid analogs ranging in polarity from ... More
Structural effects on the interaction of sterols with the (Ca2+ + Mg2+)-ATPase.
AuthorsMichelangeli F, East JM, Lee AG
JournalBiochim Biophys Acta
PubMed ID2142439
The fluorescence quenching properties of a brominated derivative of androstenol 5 alpha,6 beta-dibromoandrostan-3 beta-ol have been used to study binding to phospholipid bilayers and to the (Ca2+ + Mg2+)-ATPase purified from sarcoplasmic reticulum of rabbit skeletal muscle. It is shown that androstenol is excluded from the phospholipid/protein interface of the ... More
Membrane fluidity and oxygen diffusion in cholesterol-enriched endothelial cells.
In this study, we measured the influence of cholesterol rigidification on oxygen permeability in human endothelial cell monolayer membranes (ECs). Cholesterol-induced membrane rigidification was assessed at different membrane depths by a fluorescence polarization method with diphenyl-hexatriene (DPH) and 1-(4-trimethylamino)-6-phenylhexatriene (TMA-DPH). Fluorescence quenching by oxygen was probed in preferentially labelled membrane ... More
Fluorescence and kinetic studies of the interactions of pyrethroids with the (Ca2(+) + Mg2+)-ATPase.
AuthorsMichelangeli F, Robson MJ, East JM, Lee AG
JournalBiochim Biophys Acta
PubMed ID2145036
The fluorescence quenching properties of a series of brominated and iodinated pyrethroids have been used to study the binding of pyrethroids to the (Ca2(+) + Mg2+)-ATPase purified from skeletal muscle sarcoplasmic reticulum. It is suggested that binding at the lipid/protein interface of the ATPase is weak but that binding can ... More
Photosensitization to ultraviolet irradiation and selective killing of cells following uptake of pyrene-containing fatty acid.
AuthorsFibach E, Morand O, Gatt S
JournalJ Cell Sci
PubMed ID3793789
Cells were incubated with 12-(1-pyrene)-dodecanoic acid (P12), a long-chain fatty acid to which a pyrene ring has been attached covalently. This acid was transported across the plasma membranes of cells and subsequently incorporated into their neutral lipids and phospholipids. Irradiation of these pyrene-containing cells for short periods (0.5-4 min) with ... More
Interactions of pyrene derivatives with lipid bilayers and with (Ca2+-Mg2+)-ATPase.
AuthorsJones OT, Lee AG
JournalBiochemistry
PubMed ID3158348
The intensities of fluorescence emission for pyrene and a number of its derivatives increase on binding to lipid bilayers and to the (Ca2+-Mg2+)-ATPase purified from rabbit muscle sarcoplasmic reticulum. The effect is particularly marked for the less water-soluble derivatives. Changes in intensity for monomer and excimer emission as a function ... More
The peroxin Pex14p. cDNA cloning by functional complementation on a Chinese hamster ovary cell mutant, characterization, and functional analysis.
AuthorsShimizu N, Itoh R, Hirono Y, Otera H, Ghaedi K, Tateishi K, Tamura S, Okumoto K, Harano T, Mukai S, Fujiki Y
JournalJ Biol Chem
PubMed ID10212238
Rat cDNA encoding a 376-amino acid peroxin was isolated by functional complementation of a peroxisome-deficient Chinese hamster ovary cell mutant, ZP110, of complementation group 14 (CG14). The primary sequence showed 28 and 24% amino acid identity with the yeast Pex14p from Hansenula polymorpha and Saccharomyces cerevisiae, respectively; therefore, we termed ... More
Genetic evidence supporting the role of peroxisome assembly factor (PAF)-1 in peroxisome biogenesis. Polymerase chain reaction detection of a missense mutation in PAF-1 of Chinese hamster ovary cells.
AuthorsAllen LA, Hope L, Raetz CR, Thieringer R
JournalJ Biol Chem
PubMed ID7512952
The peroxisome/plasmalogen-deficient Chinese hamster ovary (CHO) mutant cell line ZR-78.1 contains a missense mutation in its cDNA-encoding peroxisome assembly factor-1 (PAF-1). Using a rapid polymerase chain reaction assay, we now demonstrate that the genome of ZR-78.1 contains only the mutant allele. When mutant ZR-78.1 is fused with wild-type karyoplasts, occasional ... More
Fluorescence imaging of pyrene-labeled lipids in living cells.
AuthorsTanhuanpää K, Virtanen J, Somerharju P
JournalBiochim Biophys Acta
PubMed ID10996655
Microscopic imaging of fluorescent lipid derivatives is a powerful tool to study membrane organization and lipid trafficking but it is complicated by cellular autofluorescence background and photobleaching of the fluorophore as well as by the difficulty to selectively image membranes stacked on top of each other. Here we describe protocols ... More
Pyrene derivatives as markers of transbilayer effect of lipid peroxidation on neuronal membranes.
AuthorsViani P, Cervato G, Cestaro B
JournalBiochim Biophys Acta
PubMed ID2025635
Two different pyrene derivatives, namely 12-(1-pyrene)dodecanoic acid (P12-FA) and N-(12-(1-pyrene)dodecanoyl)-galactosylsphingosine I3-sulfate (P12-CS) have been used to follow lipid peroxidation both in model and natural membranes. The malondialdehyde (MDA) production in small unilamellar vesicles of dipalmitoylphosphatidylcholine/arachidonic acid (80:20, molar ratio), symmetrically labelled with both probes determined a progressive decrease of pyrene ... More
A possible role for plasmalogens in protecting animal cells against photosensitized killing.
AuthorsZoeller RA, Morand OH, Raetz CR
JournalJ Biol Chem
PubMed ID3403547
Chinese hamster ovary (CHO) cells incorporate 12-(1'-pyrene) dodecanoic acid (P12) into membrane lipids. Exposure of P12-labeled cells to long wavelength ultraviolet light causes cell killing, presumably because excitation of the pyrene moiety (a photosensitizer) leads to the generation of reactive oxygen species. Cytotoxicity is dependent upon the concentration of P12 ... More
Fluorescence excimer formation imaging: a new technique to investigate association to cells and membrane behavior of glycolipids.
AuthorsTerzaghi A, Tettamanti G, Palestini P, Acquotti D, Bottiroli G, Masserini M
JournalEur J Cell Biol
PubMed ID7889988
A new fluorescence ratio imaging technique aiming to monitor the lateral distribution of pyrene-labeled lipids in the membranes through visualization of the excimer/monomer (E/M) intensity ratio, has been set up, studying the association of a fluorescent derivative of GM1 ganglioside (pyreneGM1) to rat cerebellar granule cells in culture. The imaging ... 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
Evidence that photodynamic stress kills Zellweger fibroblasts by a nonapoptotic mechanism.
AuthorsSpisni E, Cavazzoni M, Griffoni C, Calzolari E, Tomasi V
JournalBiochim Biophys Acta
PubMed ID9551086
Zellweger fibroblasts, which are devoid of peroxisomes and fail to synthesize plasmalogens, are very sensitive to the killing effect triggered by UV-activated 12-(1-pyrene) dodecanoic acid (P12). Although in some studied performed, it is assumed that reactive oxygen species (ROS) may damage plasma membrane causing necrosis, other studies suggest that ROS ... 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
Spectroscopical characteristics of galactosylceramide-pyrene and ceramide-pyrene incorporated in model and in clathrin coated vesicles.
AuthorsNicolas E, Lavialle F, Alfsen A
JournalChem Phys Lipids
PubMed ID8348676
In order to control the ability of two pyrene-sphingolipids (ceramide-pyrene (Cpyr) and galactosylceramide-pyrene (GCpyr)) to monitor the changes in the lipid bilayer dynamics of cellular membranes, their incorporation in three populations of clathrin coated vesicles which differ in their structural characteristics (Bomsel et al. (1988) Biochemistry 27, 6808-6812) was studied ... More
Disruption of the interaction of the longer isoform of Pex5p, Pex5pL, with Pex7p abolishes peroxisome targeting signal type 2 protein import in mammals. Study with a novel Pex5-impaired Chinese hamster ovary cell mutant.
AuthorsMatsumura T, Otera H, Fujiki Y
JournalJ Biol Chem
PubMed ID10767287
We isolated peroxisome biogenesis-defective Chinese hamster ovary cell mutants from TKaG2 cells, wild-type CHO-K1 cells transformed with two cDNAs encoding rat Pex2p and peroxisome targeting signal (PTS) type 2-tagged green fluorescent protein, by the 9-(1'-pyrene)nonanol/UV selection method. Ten mutant clones showed cytosolic PTS2-green fluorescent protein, indicative of a defect in ... More
Effects of fluorophore structure and hydrophobicity on the uptake and metabolism of fluorescent lipid analogs.
AuthorsNaylor BL, Picardo M, Homan R, Pownall HJ
JournalChem Phys Lipids
PubMed ID1934193
Cellular transport and metabolism of fatty acids are integral components of lipid metabolism, but the mechanisms and regulation involved are poorly understood. A variety of commercially available fluorescent analogs of fatty acids, are potentially useful probes for the study of lipid metabolism by such techniques as cell sorting and fluorescence ... 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
Photosensitization of human bladder carcinoma cells by pyrene-dodecanoic acid: quantitative analysis of the cytotoxicity.
AuthorsFibach E, Rachmilewitz EA, Gatt S
JournalRes Exp Med (Berl)
PubMed ID1636063
The effects of photochemotherapy with the fluorescent fatty acid pyrenedodecanoic acid (P12) and long-wavelength ultraviolet (UVA) light on cells derived from human bladder carcinoma were studied. Exposure of these anchorage-dependent cells to P12 either in monolayers of adherent cells or in suspension resulted in a time-related uptake of P12 and ... More
High-affinity binding of very-long-chain fatty acyl-CoA esters to the peroxisomal non-specific lipid-transfer protein (sterol carrier protein-2).
AuthorsDansen TB, Westerman J, Wouters FS, Wanders RJ, van Hoek A, Gadella TW, Wirtz KW
JournalBiochem J
PubMed ID10085244
Binding of fluorescent fatty acids to bovine liver non-specific lipid-transfer protein (nsL-TP) was assessed by measuring fluorescence resonance energy transfer (FRET) between the single tryptophan residue of nsL-TP and the fluorophore. Upon addition of pyrene dodecanoic acid (Pyr-C12) and cis-parinaric acid to nsL-TP, FRET was observed indicating that these fatty ... More
Spectrofluorometric measurements of the dispersion state of pyrenedodecanoic acid and its uptake by cultured cells and liposomes.
AuthorsGatt S, Fibach E
JournalBiochim Biophys Acta
PubMed ID3166380
Pyrene dodecanoic acid (P12), a medium-chain fatty acid to which the fluorescent probe pyrene is covalently linked, showed a considerable increase in fluorescence when the probe was introduced into a hydrophobic environment. Also, when closely packed in an aggregate, an energy transfer between two adjacent molecules of pyrene occurred, resulting ... More
Pyrene butanol--an efficient, selective and non-metabolized photosensitizing agent for human myeloid leukemia cells.
AuthorsFibach E, Rabia SA, Gatt S
JournalLeuk Res
PubMed ID1625470
Methods for ex vivo purging of neoplastic cells from harvested marrow are being developed to increase the efficacy of autologous transplantation. One approach is selective photosensitization, using sensitizing compounds and light radiation. Pyrene-containing fatty acids and lipids are potent photosensitizers, e.g. 12-(1-pyrene)dodecanoic acid (P12), is taken up preferentially by leukemic ... More
Synthesis of pyrene derivatives of cerebroside sulfate and their use for determining arylsulfatase A activity.
AuthorsMarchesini S, Viani P, Cestaro B, Gatt S
JournalBiochim Biophys Acta
PubMed ID2564282
Two fluorescent derivatives of cerebroside sulfate ('sulfatide') have been synthesized and used as substrates for determining arylsulfatase A activity. These were 12-(1-pyrene)dodecanoyl cerebroside sulfate (P12-sulfatide) and 12(1-pyrenesulfonylamido)dodecanoyl cerebroside sulfate (PSA12-sulfatide). When incubated at pH 5.0 in the presence of 5 mM MnCl2 and 5.5 mM of taurodeoxycholate, either substrate was ... More
Uptake and degradation of several pyrenesphingomyelins by skin fibroblasts from control subjects and patients with Niemann-Pick disease. Effect of the structure of the fluorescent fatty acyl residue.
AuthorsLevade T, Gatt S, Salvayre R
JournalBiochem J
PubMed ID2018477
Three fluorescent analogues of sphingomyelin (SPM), each containing pyrene in the fatty acyl residue, were synthesized and employed for the study of their mode of uptake by, and degradation within, intact cultured human skin fibroblasts. These were prepared by condensing sphingosylphosphocholine and the following fatty acids: pyrenedodecanoic acid (P12), pyrenesulphonylaminoundecanoic ... 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
Use of the fluorescence activated cell sorter for studying uptake of fluorescent fatty acids into cultured cells.
AuthorsNahas N, Fibach E, Giloh H, Gatt S
JournalBiochim Biophys Acta
PubMed ID3466651
A procedure is described for the continuous monitoring and recording of fatty acid transport into cultured cells. Uptake of the fluorescent fatty acid derivative, 12-(1-pyrene)dodecanoic acid [P12] by a suspension of human promyelocytic leukemia (HL-60) cells was analyzed and recorded by the Fluorescence Activated Cell Sorter (FACS). A major advantage ... More
A milling crowd model for local and long-range obstructed lateral diffusion. Mobility of excimeric probes in the membrane of intact erythrocytes.
AuthorsEisinger J, Flores J, Petersen WP
JournalBiophys J
PubMed ID3778578
A new model for lateral diffusion, the milling crowd model (MC), is proposed and is used to derive the dependence of the monomeric and excimeric fluorescence yields of excimeric membrane probes on their concentration. According to the MC model, probes migrate by performing spatial exchanges with a randomly chosen nearest ... More