High spatial resolution confocal microscopy of young MDCK cells stained with the lipophilic probe 1,1'-dihexadecyl-3,3,3',3'- tetramethylindocarbocyanine perchlorate (DiIC(16)) revealed a reticulated fluorescence pattern on the apical membrane. DiIC(16) was delivered as crystals to live cells to minimize possible solvent perturbations of the membrane lipids. The ratio of the integrated fluorescence ... More
Induced circular dichroism of incorporated fluorescent cholesteryl esters and polar lipids as a probe of human serum low density lipoprotein structure and melting.
AuthorsSklar LA, Craig IF, Pownall HJ
JournalJ Biol Chem
PubMed ID7217083
Two fluorescent cholesteryl esters, cholesteryl cis-parinarate and cholestatrienyl oleate, have been incorporated into human serum low density lipoprotein (LDL) by two alternative procedures. The core location of the incorporated cholesterol esters and the surface location of cis-parinaric acid and cholestatrienol are verified by fluorescence energy transfer. In these LDL preparations, ... More
Resonance energy transfer between guanine nucleotide binding protein subunits and membrane lipids.
AuthorsRemmers AE, Neubig RR
JournalBiochemistry
PubMed ID7680231
'Resonance energy transfer was used to estimate the distances of closest approach between fluorescent labels on G protein subunits (alpha, beta, and gamma) and the phospholipid bilayer surface. Fluorescein-labeled alpha, beta, and gamma subunits were the energy transfer donors and hexadecylaminoeosin (HAE) in phospholipid vesicles was the acceptor. Bovine brain ... More
Location of the epidermal growth factor binding site on the EGF receptor. A resonance energy transfer study.
AuthorsCarraway KL, Koland JG, Cerione RA
JournalBiochemistry
PubMed ID2271554
'As a first step toward developing a structural map of key sites on the epidermal growth factor (EGF) receptor, we have used resonance energy transfer to measure the distance of closest approach between the receptor-bound growth factor molecule and lipid molecules at the surface of the plasma membrane. EGF, specifically ... More
Kinetics of Cdc42 membrane extraction by Rho-GDI monitored by real-time fluorescence resonance energy transfer.
AuthorsNomanbhoy TK, Erickson JW, Cerione RA
JournalBiochemistry
PubMed ID10026253
'The mechanisms underlying the ability of the Rho-GDP dissociation inhibitor (RhoGDI) to elicit the release of Rho-related GTP-binding proteins from membranes is currently unknown. In this report, we have set out to address this issue by using fluorescence resonance energy transfer approaches to examine the functional interactions of the RhoGDI ... More
The location of fluorescence probes with charged groups in model membranes.
AuthorsKachel K, Asuncion-Punzalan E, London E
JournalBiochim Biophys Acta
PubMed ID9814853
'The location of commonly used charged fluorescent membrane probes in membranes was determined in order to: (1) investigate the relationship between the structure of hydrophobic molecules and their depth within membranes; and (2) aid interpretation of experiments in which these fluorescent probes are used to examine membrane structure. Membrane depth ... More
Novel fluorescein-based flow-cytometric method for detection of lipid peroxidation.
AuthorsMakrigiorgos GM, Kassis AI, Mahmood A, Bump EA, Savvides P
JournalFree Radic Biol Med
PubMed ID8958133
'The novel property of fluorescein to detect peroxyl radicals is demonstrated. On the basis of this observation, a fluorescein-based, flow-cytometric method to directly and continuously detect free radicals generated in cell membranes during lipid peroxidation has been developed. 5- and 6-Carboxyfluorescein (5-/6-CF) free in solution and fluorescein-labeled polylysine lose their ... More
Insulin effect on translational diffusion of lipids and proteins in the plasma membrane of isolated rat hepatocytes.
AuthorsStuschke M, Bojar H
JournalBiochim Biophys Acta
PubMed ID3890961
'The effects of insulin (10(-10)-10(-8) mol/l) on lateral diffusion of three fluorescent lipid probes, 1-acyl-2-(N-4-nitrobenzo-2-oxa-1,3-diazole)aminocaproyl phosphatidylcholine (NBD-PC), 5-(N-hexadecanoyl)aminofluorescein (F-C16), 5-(N-dodecanoyl)aminofluorescein (F-C12), and of fluorescein isothiocyanate-labeled proteins in the plasma membrane of intact rat hepatocytes were studied by the technique of fluorescence recovery after photobleaching. The absolute lateral diffusion coefficients of ... More
Insertion of fluorescent fatty acid probes into the outer membranes of the pathogenic spirochaetes Treponema pallidum and Borrelia burgdorferi.
AuthorsCox DL, Radolf JD
JournalMicrobiology
PubMed ID11320119
'The authors examined the ability of octadecanoyl (C(18)), hexadecanoyl (C(16)) and dodecanoyl (C(12)) fatty acid (FA) conjugates of 5-aminofluorescein (OAF, HAF and DAF, respectively) to insert into the outer membranes (OMs) of Treponema pallidum, Borrelia burgdorferi and Escherichia coli. Biophysical studies have demonstrated that these compounds stably insert into phospholipid ... More
Delayed "all-or-none" activation of inositol 1,4,5-trisphosphate-dependent calcium signaling in single rat hepatocytes.
AuthorsChiavaroli C, Bird G, Putney JW
JournalJ Biol Chem
PubMed ID7523388
'When single rat hepatocytes were stimulated with the phospholipase C-activating hormone, vasopressin (from 300 pM to 1 microM), the [Ca2+]i signals were always "all-or-none" responses. At low concentrations of vasopressin, Ca2+ release was maximal because liberation of additional inositol 1,4,5-trisphosphate (IP3) by photolysis of its caged precursor at the top ... 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
Fluorescence recovery after photobleaching (FRAP) experiments under conditions of uniform disk illumination. Critical comparison of analytical solutions, and a new mathematical method for calculation of diffusion coefficient D.
AuthorsLopez A, Dupou L, Altibelli A, Trotard J, Tocanne JF
JournalBiophys J
PubMed ID3395663
'A simple fluorescence recovery after photobleaching (FRAP) apparatus using a fluorescence microscope with a conventional mercury arc lamp, working under conditions of "uniform disk illumination" is described. This set-up was designed essentially for the use of anthracene as fluorescent probe, which is bleached (photodimerization reaction) by illumination in the near ... More
The use of fluorescein-dipalmitoylphosphatidylethanolamine for measuring pH-changes in the internal compartment of phospholipid vesicles.
AuthorsThelen M, Petrone G, O'Shea PS, Azzi A
JournalBiochim Biophys Acta
PubMed ID6743649
'The synthesis and characterisation of fluorescein-phosphatidylethanolamine (FPE) is described. The effects of dielectric constant, ionic strength and ambient pH upon the optical absorbance properties of FPE are presented. It is shown that under appropriate conditions, FPE rapidly and quantitatively reports the pH of the aqueous bulk phases when incorporated into ... More
Structural mapping of Fc receptor bound immunoglobulin E: proximity to the membrane surface of the antibody combining site and another site in the Fab segments.
AuthorsBaird B, Holowka D
JournalBiochemistry
PubMed ID4084517
'Resonance energy-transfer methods have been used to investigate the structure of immunoglobulin E (IgE) bound to its high-affinity receptor on plasma membrane vesicles derived from rat basophilic leukemia cells. The structural mapping of receptor-bound IgE was initiated in an earlier study [Holowka, D., & Baird, B. (1983) Biochemistry 22, 3475], ... More
Fluorescent cholesteryl esters in the core of low density lipoprotein.
AuthorsSklar LA, Mantulin WW, Pownall HJ
JournalBiochem Biophys Res Commun
PubMed ID7092879
Structural investigation of reconstituted chloroplast ATPase with fluorescence measurements.
AuthorsBaird BA, Pick U, Hammes GG
JournalJ Biol Chem
PubMed ID155694
Lateral diffusion of lipids in model and natural membranes.
AuthorsTocanne JF, Dupou-Cézanne L, Lopez A
JournalProg Lipid Res
PubMed ID8022844
Comparative lateral diffusion of fluorescent lipid analogues in phospholipid multibilayers.
AuthorsDerzko Z, Jacobson K
JournalBiochemistry
PubMed ID7470449
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
Spectral properties of fluorescent dyes in lecithin vesicles. Probes for the structure of lipid bilayer membranes and for membrane potentials.
AuthorsPohl GW
JournalZ Naturforsch [C]
PubMed ID136128
Three fluorescent dyes (a merocyanin, a cyanin and a modified umbeliferon) have been incorporated into vesicles from three different lecithins. Their absorption spectrum, emission spectrum and fluorescence polarization in the lipid vesicles has been studied and compared with the spectral properties in homogeneous organic solutions. The reactivity of lipid bound ... More
The impenetrability of 5-(N-hexadecanoyl)aminofluoroscein through endothelial cell monolayers is dependent upon its solution properties, not the presence of tight junctions.
AuthorsStolz DB, Mahoney MG, Jacobson BS
JournalBiochem Biophys Res Commun
PubMed ID1567423
The solution properties of two fluorescent lipophilic analogues were examined in conjunction with their ability to penetrate the tight junctions of bovine aortic endothelial cell monolayers. 5-(N-dodecanoyl)aminofluoroscein was shown to label both the apical and basolateral plasma membrane domains of confluent monolayers at 4 degrees C and pH 7.3, but ... More
Extracellular pH dynamics of retinal horizontal cells examined using electrochemical and fluorometric methods.
Extracellular H(+) has been hypothesized to mediate feedback inhibition from horizontal cells onto vertebrate photoreceptors. According to this hypothesis, depolarization of horizontal cells should induce extracellular acidification adjacent to the cell membrane. Experiments testing this hypothesis have produced conflicting results. Studies examining carp and goldfish horizontal cells loaded with the ... More
Depolarization of isolated horizontal cells of fish acidifies their immediate surrounding by activating V-ATPase.
AuthorsJouhou H, Yamamoto K, Homma A, Hara M, Kaneko A, Yamada M,
JournalJ Physiol
PubMed ID17932147
In order to interpret the formation of receptive field surrounds in retinal neurons, a proton-mediated mechanism was proposed to mediate feedback from horizontal cells (HCs) to cone photoreceptors. To verify the idea that depolarized HCs release protons, we measured, by a fluorescence ratiometric technique, the pH of the immediate external ... More
Fluorescence measurements of serotonin-induced V-ATPase-dependent pH changes at the luminal surface in salivary glands of the blowfly Calliphora vicina.
AuthorsRein J, Zimmermann B, Hille C, Lang I, Walz B, Baumann O,
JournalJ Exp Biol
PubMed ID16621952
Secretion in blowfly salivary glands is induced by the neurohormone serotonin and powered by a vacuolar-type H(+)-ATPase (V-ATPase) located in the apical membrane of the secretory cells. We have established a microfluorometric method for analysing pH changes at the luminal surface of the secretory epithelial cells by using the fluorescent ... More
How curved membranes recruit amphipathic helices and protein anchoring motifs.
Lipids and several specialized proteins are thought to be able to sense the curvature of membranes (MC). Here we used quantitative fluorescence microscopy to measure curvature-selective binding of amphipathic motifs on single liposomes 50-700 nm in diameter. Our results revealed that sensing is predominantly mediated by a higher density of ... More
Serum lipoprotein structure: resonance energy transfer localization of fluorescent lipid probes.
AuthorsSklar LA, Doody MC, Gotto AM, Pownall HJ
JournalBiochemistry
PubMed ID7387988
The location of several fluorescent chromophores in lipoproteins has been determined by using resonance energy transfer. The primary acceptor is 5-(N-hexadecanoylamino)fluorescein whose chromophore is shown to reside at the lipoprotein surface at pH 7.4. Polar donors include cis-parinaric acid (cis,trans,trans,cis-9,11,13,15-octadecatetraenoic acid), trans-parinaric acid (all-trans-9,11,13,15-octadecatetraenoic acid), and 16-(9-anthroyloxy)palmitic acid; nonpolar donors ... More
Incorporation of defined cholesteryl esters into lipoproteins using cholesteryl ester-rich microemulsions.
AuthorsCraig IF, Via DP, Sherrill BC, Sklar LA, Mantulin WW, Gotto AM, Smith LC
JournalJ Biol Chem
PubMed ID7053374
Cholesteryl ester transfer to low density lipoproteins (LDL) was achieved either by incubation of a cholesteryl ester-rich lipid microemulsion with whole plasma or by incubation of the cholesteryl ester-rich microemulsion with lipoprotein-free plasma and LDL. Sequential ultracentrifugal isolation gave LDL that contained between 5 and 37% of the cholesteryl ester ... More
Restricted photorelease of biologically active molecules near the plasma membrane.
AuthorsSuga T, Hirano M, Takayanagi M, Koshimoto H, Watanabe A
JournalBiochem Biophys Res Commun
PubMed ID9878552
An evanescent wave of ultraviolet light was successfully used to release biologically active molecules from caged compounds in living cells. The evanescent wave was generated by the total internal reflection in a limited region near the plasma membrane attached to the illuminated interface. At first, the photolysis efficiency of the ... More
Transverse distance between the membrane and the agonist binding sites on the Torpedo acetylcholine receptor: a fluorescence study.
AuthorsValenzuela CF, Weign P, Yguerabide J, Johnson DA
JournalBiophys J
PubMed ID8011898
Fluorescence dipolar resonance energy transfer between a receptor-bound fluorescent agonist, dansyl-C6-choline, and two membrane-partitioned fluorescent probes, C18-rhodamine and C12-eosin, was used to measure the transverse distance between the acetylcholine (ACh) binding sites on the intact Torpedo nicotinic acetylcholine receptor (nAChR) and the surface of the lipid membrane. Control experiments demonstrated ... More
C11-BODIPY(581/591), an oxidation-sensitive fluorescent lipid peroxidation probe: (micro)spectroscopic characterization and validation of methodology.
AuthorsDrummen GP, van Liebergen LC, Op den Kamp JA, Post JA
JournalFree Radic Biol Med
PubMed ID12160930
C11-BODIPY(581/591) is a fluorescent radio-probe for indexing lipid peroxidation and antioxidant efficacy in model membrane systems and living cells, with excellent characteristics: (i) emission in the visible range of the electromagnetic spectrum, with good spectral separation of the nonoxidized (595 nm) and oxidized (520 nm) forms; (ii) has a high ... More
Alterations in lateral lipid mobility in the plasma membrane of urodelean ectodermal cells during gastrulation.
AuthorsDupou L, Gualandris L, Lopez A, Duprat AM, Tocanne JF
JournalExp Cell Res
PubMed ID3556431
The mobility characteristics of lipids were studied in the plasmalemma of dissociated presumptive ectodermal cells from embryos of Pleurodeles Waltl at different stages of development, from early blastula to early neurula, using a Fluorescence Recovery After Photobleaching technique (FRAP), after incorporation of the lipophilic fluorescent probe 5N-(hexadecanoyl)-aminofluoresceine (HEDAF) into the ... More
The lateral diffusion of lipid probes in the surface membrane of Schistosoma mansoni.
AuthorsFoley M, MacGregor AN, Kusel JR, Garland PB, Downie T, Moore I
JournalJ Cell Biol
PubMed ID3745270
The technique of fluorescence recovery after photobleaching was used to measure the lateral diffusion of fluorescent lipid analogues in the surface membrane of Schistosoma mansoni. Our data reveal that although some lipids could diffuse freely others exhibited restricted lateral diffusion. Quenching of lipid fluorescence by a non-permeant quencher, trypan blue, ... More
A simple and sensitive flow cytometric assay for the determination of the cytotoxic activity of human natural killer cells.
AuthorsRadosevic K, Garritsen HS, Van Graft M, De Grooth BG, Greve J
JournalJ Immunol Methods
PubMed ID2273268
A new, simple and sensitive flow cytometric assay for the determination of the cytotoxic activity of human natural killer cells is described. The assay is based on the use of two fluorochromes. The target cell population is stained with one fluorochrome (octadecylamine-fluorescein isothiocyanate, F-18) prior to incubation with the effector ... More
Structural studies on the membrane-bound immunoglobulin E-receptor complex. 1. Characterization of large plasma membrane vesicles from rat basophilic leukemia cells and insertion of amphipathic fluorescent probes.
AuthorsHolowka D, Baird B
JournalBiochemistry
PubMed ID6225455
In order to investigate the properties of the membrane-bound IgE-receptor complex, a simple procedure has been adapted for preparing large plasma membrane vesicles from rat basophilic leukemia cells. These vesicles pinch off from the adherent cells after treatment with 2 mM N-ethylmaleimide or 50 mM formaldehyde and 1 mM dithiothreitol, ... More
The lateral mobility of the (Na+,K+)-dependent ATPase in Madin-Darby canine kidney cells.
AuthorsJesaitis AJ, Yguerabide J
JournalJ Cell Biol
PubMed ID3007531
Fluorescence microphotolysis (recovery after photobleaching) was used to determine the lateral mobility of the (Na+,K+)ATPase and a fluorescent lipid analogue in the plasma membrane of Madin-Darby canine kidney (MDCK) cells at different stages of development. Fluorescein-conjugated Fab' fragments prepared from rabbit anti-dog (Na+,K+)ATPase antibodies (IgG) and 5-(N-hexadecanoyl)aminofluorescein (HEDAF) were used ... More
Temperature-dependent lateral and transverse distribution of the epidermal growth factor receptor in A431 plasma membranes.
AuthorsAzevedeo JR, Johnson DA
JournalJ Membr Biol
PubMed ID2077131
To elucidate further the structure and molecular dynamics of the epidermal growth factor receptor, temperature-dependent aggregation and extracellular protrusion of the epidermal growth factor receptor in isolated plasma membranes from A431 cells were examined by fluorescence energy-transfer techniques. Epidermal growth factor was labeled at the amino terminus with either fluorescein ... More
Maintenance and regulation of the pH microclimate at the luminal surface of the distal colon of guinea-pig.
AuthorsGenz AK, v Engelhardt W, Busche R
JournalJ Physiol
PubMed ID10332098
1. The fluorescent dye 5-N-hexadecanoyl-aminofluorescein (HAF) was used to study the mechanisms involved in maintaining a relatively constant luminal surface pH (pHs) in the distal colon of the guinea-pig. The fatty acyl chain of the HAF molecule inserts into the apical membrane of epithelial cells. This allows a continuous measurement ... More
Comparative study of the lateral motion of extrinsic probes and anthracene-labelled constitutive phospholipids in the plasma membrane of Chinese hamster ovary cells.
AuthorsDupou L, Lopez A, Tocanne JF
JournalEur J Biochem
PubMed ID3345753
9-(2-Anthryl)-nonanoic acid is a new photoactivatable fluorescent probe which has been designed for the study of the lateral diffusion and distribution of lipids in biological membranes by means of the anthracene photodimerization reaction. This anthracene fatty acid can be incorporated metabolically into the glycerophospholipids (phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol) of Chinese ... More
Effect of cell substratum on lateral mobility of lipids in the plasma membrane of vascular endothelial cells.
AuthorsTournier JF, Lopez A, Tocanne JF
JournalExp Cell Res
PubMed ID2917598
Bovine vascular endothelial cells can be maintained in a highly differentiated state in vitro, either by the addition of fibroblast growth factor (FGF) to the culture medium or by plating the cells on extracellular matrix (ECM)-coated dishes. Under these conditions the cells proliferate actively and at confluence form a tightly ... More
Antibody-induced conformational changes in the Torpedo nicotinic acetylcholine receptor: a fluorescence study.
AuthorsValenzuela CF, Dowding AJ, Arias HR, Johnson DA
JournalBiochemistry
PubMed ID8204595
Quantitative fluorescence spectroscopy was used to develop a structural picture of the effects of two monoclonal antibodies (mAbs) on the conformation of the Torpedo nicotinic acetylcholine receptor (nAChR). The two mAbs (A6 and B1) examined selectively blocked ligand binding to either the high-affinity (A) or the low-affinity (B) binding sites ... More
Resonance energy transfer study of membrane-bound aggregates of the sarcoplasmic reticulum calcium ATPase.
AuthorsFagan MH, Dewey TG
JournalJ Biol Chem
PubMed ID2936737
The aggregation of the membrane-bound calcium ATPase from sarcoplasmic reticulum has been studied by resonance energy transfer. The temperature dependence of resonance energy transfer from a fluorescent membrane lipid donor to an acceptor covalently linked to the Ca2+ ATPase was observed for the native sarcoplasmic reticulum vesicles and for purified ... More
Differences in the transbilayer and lateral motions of fluorescent analogs of phosphatidylcholine and phosphatidylethanolamine in the apical plasma membrane of bovine aortic endothelial cells.
AuthorsJulien M, Tournier JF, Tocanne JF
JournalExp Cell Res
PubMed ID8375469
In the plasma membrane of various eucaryotic cell types, in particular blood platelets and erythrocytes, it is known that phospholipids are asymmetrically distributed between the two leaflets of the lipid bilayer and that this transverse asymmetry is controlled by an aminophospholipid translocase activity. In this respect, it was of interest ... More
The lateral motion of lipid molecules in the apical plasma membrane of endothelial cells is reversibly affected by the presence of cell junctions.
AuthorsTournier JF, Lopez A, Gas N, Tocanne JF
JournalExp Cell Res
PubMed ID2924796
As with other epithelia, the question arises of whether the endothelial cell junctions participate in cell polarization, restrict the localization of lipid molecules, and lead to differences in their lateral motion between the apical and basolateral plasma membranes. We found that in bovine aortic endothelium in culture, the localization of ... More
High lateral mobility of endogenous and transfected alkaline phosphatase: a phosphatidylinositol-anchored membrane protein.
AuthorsNoda M, Yoon K, Rodan GA, Koppel DE
JournalJ Cell Biol
PubMed ID2889741
The lateral mobility of alkaline phosphatase (AP) in the plasma membrane of osteoblastic and nonosteoblastic cells was estimated by fluorescence redistribution after photobleaching in embryonic and in tumor cells, in cells that express AP naturally, and in cells transfected with an expression vector containing AP cDNA. The diffusion coefficient (D) ... More
Continuous fluorescence microphotolysis of anthracene-labeled phospholipids in membranes. Theoretical approach of the simultaneous determination of their photodimerization and lateral diffusion rates.
AuthorsFerrières X, Lopez A, Altibelli A, Dupou-Cezanne L, Lagouanelle JL, Tocanne JF
JournalBiophys J
PubMed ID2765646
Anthracene is a fluorescent and photoactivatable (dimerization) group which can be used for investigating the lateral distribution and dynamics of lipids in membranes. In fluorescence recovery after photobleaching or in microphotolysis experiments, and when using this fluorophore, the bleaching (or microphotolysis) step in the illuminated part of the membrane is ... More
Lipid domains of mycobacteria studied with fluorescent molecular probes.
AuthorsChristensen H, Garton NJ, Horobin RW, Minnikin DE, Barer MR
JournalMol Microbiol
PubMed ID10200973
The complex mycobacterial cell envelope is recognized as a critical factor in our failure to control tuberculosis, leprosy and other non-tuberculous pathogens. Although its composition has been extensively determined, many details regarding the organization of the envelope remain uncertain. This is particularly so for the non-covalently bound lipids, whose natural ... More