Structure refinement and membrane positioning of selectively labeled OmpX in phospholipid nanodiscs

Energy Technology Data Exchange (ETDEWEB)

Hagn, Franz, E-mail: franz.hagn@tum.de; Wagner, Gerhard, E-mail: gerhard-wagner@hms.harvard.edu [Harvard Medical School, Department of Biological Chemistry and Molecular Pharmacology (United States)

2015-04-15

NMR structural studies on membrane proteins are often complicated by their large size, taking into account the contribution of the membrane mimetic. Therefore, classical resonance assignment approaches often fail. The large size of phospholipid nanodiscs, a detergent-free phospholipid bilayer mimetic, prevented their use in high-resolution solution-state NMR spectroscopy so far. We recently introduced smaller nanodiscs that are suitable for NMR structure determination. However, side-chain assignments of a membrane protein in nanodiscs still remain elusive. Here, we utilized a NOE-based approach to assign (stereo-) specifically labeled Ile, Leu, Val and Ala methyl labeled and uniformly {sup 15}N-Phe and {sup 15}N-Tyr labeled OmpX and calculated a refined high-resolution structure. In addition, we were able to obtain residual dipolar couplings (RDCs) of OmpX in nanodiscs using Pf1 phage medium for the induction of weak alignment. Back-calculated NOESY spectra of the obtained NMR structures were compared to experimental NOESYs in order to validate the quality of these structures. We further used NOE information between protonated lipid head groups and side-chain methyls to determine the position of OmpX in the phospholipid bilayer. These data were verified by paramagnetic relaxation enhancement (PRE) experiments obtained with Gd{sup 3+}-modified lipids. Taken together, this study emphasizes the need for the (stereo-) specific labeling of membrane proteins in a highly deuterated background for high-resolution structure determination, particularly in large membrane mimicking systems like phospholipid nanodiscs. Structure validation by NOESY back-calculation will be helpful for the structure determination and validation of membrane proteins where NOE assignment is often difficult. The use of protein to lipid NOEs will be beneficial for the positioning of a membrane protein in the lipid bilayer without the need for preparing multiple protein samples.

Insulin stimulation of phospholipid methylation in isolated rat adipocyte plasma membranes.

OpenAIRE

Kelly, K L; Kiechle, F L; Jarett, L

1984-01-01

Partially purified plasma membranes prepared from rat adipocytes contain N-methyltransferase(s) that utilize(s) S-adenosyl-L-methionine to synthesize phosphatidylcholine from phosphatidylethanolamine. The incorporation of [3H]methyl from S-adenosyl-L-[methyl-3H]methionine into plasma membrane phospholipids was linear with incubation time and plasma membrane protein concentration and was inhibited in a dose-dependent manner by both S-adenosyl-L-homocysteine and 3-deazadenosine. The addition of...

Interaction of abscisic acid with phospholipid membranes

International Nuclear Information System (INIS)

Stillwell, W.; Brengle, B.; Hester, P.; Wassall, S.T.

1989-01-01

The plant hormone abscisic acid (ABA) is shown, under certain conditions, to greatly enhance the permeability of phospholipid bilayer membranes to the nonelectrolyte erythritol (followed spectrophotometrically by osmotic swelling) and the anion carboxyfluorescein (followed by fluorescence). The hormone is ineffective with single- and mixed-component phosphatidylcholine membranes in the liquid-crystalline or gel states. In contrast, substantial ABA-induced permeability is measured for two-component membranes containing lipids with different polar head groups or containing phosphatidylcholines with different acyl chains at temperatures where gel and liquid-crystalline phases coexist. Despite the large ABA-induced enhancement in bilayer permeability, no evidence for a substantial change at the molecular level was seen in the membranes by magnetic resonance techniques. 13 C NMR spin-lattice relaxation times, T 1 , in sonicated unilamellar vesicles and ESR of spin-labeled fatty acids intercalated into membranes showed negligible effect on acyl chain order and dynamics within the bilayer, while 31 P NMR of sonicated unilamellar vesicles indicated negligible effect on molecular motion and conformation in the head-group region. The authors propose that, instead of causing a general nonspecific perturbation to the membrane, the hormone acts at membrane defects formed due to mismatch in molecular packing where two different head groups or acyl chain states interface. Increased membrane disruption by ABA at these points of membrane instability could then produce an enhancement in permeability

Checks and balances in membrane phospholipid class and acyl chain homeostasis, the yeast perspective

NARCIS (Netherlands)

de Kroon, A.I.P.M.|info:eu-repo/dai/nl/084765283; Rijken, P.J.|info:eu-repo/dai/nl/32716297X; De Smet, C.H.|info:eu-repo/dai/nl/304824224

2013-01-01

Glycerophospholipids are the most abundant membrane lipid constituents in most eukaryotic cells. As a consequence, phospholipid class and acyl chain homeostasis are crucial for maintaining optimal physical properties of membranes that in turn are crucial for membrane function. The topic of this

Biophysical properties of membrane lipids of anammox bacteria : I. Ladderane phospholipids form highly organized fluid membranes

NARCIS (Netherlands)

Boumann, Henry A.; Longo, Marjorie L.; Stroeve, Pieter; Poolman, Bert; Hopmans, Ellen C.; Stuart, Marc C. A.; Damste, Jaap S. Sinninghe; Schouten, Stefan

Anammox bacteria that are capable of anaerobically oxidizing ammonium (anammox) with nitrite to nitrogen gas produce unique membrane phospholipids that comprise hydrocarbon chains with three or five linearly condensed cyclobutane rings. To gain insight into the biophysical properties of these

Rooster sperm plasma membrane protein and phospholipid organization and reorganization attributed to cooling and cryopreservation

Science.gov (United States)

Cholesterol to phospholipid ratio is used as a representation for membrane fluidity, and predictor of cryopreservation success but results are not consistent across species and ignore the impact of membrane proteins. Therefore, this research explored the modulation of membrane fluidity and protein ...

Employment of Voltammetry in Studies of Transport Processes across Artificial Phospholipid Membranes

Czech Academy of Sciences Publication Activity Database

Šestáková, Ivana; Navrátil, Tomáš; Josypčuk, Bohdan

2016-01-01

Roč. 28, č. 11 (2016), s. 2754-2759 ISSN 1040-0397 Institutional support: RVO:61388955 Keywords : phospholipid membrane * cadmium * calcium ionophore (calcimycin) Subject RIV: CG - Electrochemistry Impact factor: 2.851, year: 2016

Kinase Associated-1 Domains Drive MARK/PAR1 Kinases to Membrane Targets by Binding Acidic Phospholipids

Energy Technology Data Exchange (ETDEWEB)

Moravcevic, Katarina; Mendrola, Jeannine M.; Schmitz, Karl R.; Wang, Yu-Hsiu; Slochower, David; Janmey, Paul A.; Lemmon, Mark A. (UPENN-MED)

2011-09-28

Phospholipid-binding modules such as PH, C1, and C2 domains play crucial roles in location-dependent regulation of many protein kinases. Here, we identify the KA1 domain (kinase associated-1 domain), found at the C terminus of yeast septin-associated kinases (Kcc4p, Gin4p, and Hsl1p) and human MARK/PAR1 kinases, as a membrane association domain that binds acidic phospholipids. Membrane localization of isolated KA1 domains depends on phosphatidylserine. Using X-ray crystallography, we identified a structurally conserved binding site for anionic phospholipids in KA1 domains from Kcc4p and MARK1. Mutating this site impairs membrane association of both KA1 domains and intact proteins and reveals the importance of phosphatidylserine for bud neck localization of yeast Kcc4p. Our data suggest that KA1 domains contribute to coincidence detection, allowing kinases to bind other regulators (such as septins) only at the membrane surface. These findings have important implications for understanding MARK/PAR1 kinases, which are implicated in Alzheimer's disease, cancer, and autism.

Correlated volume-energy fluctuations of phospholipid membranes: A simulation study

DEFF Research Database (Denmark)

Pedersen, Ulf. R.; Peters, Günther H.J.; Schröder, Thomas B.

2010-01-01

This paper reports all-atom computer simulations of five phospholipid membranes (DMPC, DPPC, DMPG, DMPS, and DMPSH) with focus on the thermal equilibrium fluctuations of volume, energy, area, thickness, and chain order. At constant temperature and pressure, volume and energy exhibit strong...... membranes, showing a similar picture. The cause of the observed strong correlations is identified by splitting volume and energy into contributions from tails, heads, and water, and showing that the slow volume−energy fluctuations derive from van der Waals interactions of the tail region; they are thus...

Bacillus subtilis alters the proportion of major membrane phospholipids in response to surfactin exposure.

Science.gov (United States)

Uttlová, Petra; Pinkas, Dominik; Bechyňková, Olga; Fišer, Radovan; Svobodová, Jaroslava; Seydlová, Gabriela

2016-12-01

Surfactin, an anionic lipopeptide produced by Bacillus subtilis, is an antimicrobial that targets the cytoplasmic membrane. Nowadays it appears increasingly apparent that the mechanism of resistance against these types of antibiotics consists of target site modification. This prompted us to investigate whether the surfactin non-producing strain B. subtilis 168 changes its membrane composition in response to a sublethal surfactin concentration. Here we show that the exposure of B. subtilis to surfactin at concentrations of 350 and 650 μg/ml (designated as SF350 and SF650, respectively) leads to a concentration-dependent growth arrest followed by regrowth with an altered growth rate. Analysis of the membrane lipid composition revealed modifications both in the polar head group and the fatty acid region. The presence of either surfactin concentration resulted in a reduction in the content of the major membrane phospholipid phosphatidylglycerol (PG) and increase in phosphatidylethanolamine (PE), which was accompanied by elevated levels of phosphatidic acid (PA) in SF350 cultures. The fatty acid analysis of SF350 cells showed a marked increase in non-branched high-melting fatty acids, which lowered the fluidity of the membrane interior measured as the steady-state fluorescence anisotropy of DPH. The liposome leakage of carboxyfluorescein-loaded vesicles resembling the phospholipid composition of surfactin-adapted cells showed that the susceptibility to surfactin-induced leakage is strongly reduced when the PG/PE ratio decreases and/or PA is included in the target bilayer. We concluded that the modifications of the phospholipid content of B. subtilis cells might provide a self-tolerance of the membrane active surfactin. Copyright © 2016 Elsevier B.V. All rights reserved.

Sequestration of polyunsaturated fatty acids in membrane phospholipids of Caenorhabditis elegans dauer larva attenuates eicosanoid biosynthesis for prolonged survival

Directory of Open Access Journals (Sweden)

Sin Man Lam

2017-08-01

Full Text Available Mechanistic basis governing the extreme longevity and developmental quiescence of dauer juvenile, a “non-ageing” developmental variant of Caenorhabditis elegans, has remained largely obscure. Using a lipidomic approach comprising multiple reaction monitoring transitions specific to distinct fatty acyl moieties, we demonstrated that in comparison to other developmental stages, the membrane phospholipids of dauer larva contain a unique enrichment of polyunsaturated fatty acids (PUFAs. Esterified PUFAs in phospholipids exhibited temporal accumulation throughout the course of dauer endurance, followed by sharp reductions prior to termination of diapause. Reductions in esterified PUFAs were accompanied by concomitant increases in unbound PUFAs, as well as their corresponding downstream oxidized derivatives (i.e. eicosanoids. Global phospholipidomics has unveiled that PUFA sequestration in membrane phospholipids denotes an essential aspect of dauer dormancy, principally via suppression of eicosanoid production; and a failure to upkeep membrane lipid homeostasis is associated with termination of dauer endurance. Keywords: Dauer larva, Phospholipids, Polyunsaturated fatty acids, Eicosanoids, Lipidomics, Caenorhabditis elegans

Lipid chain saturation and the cholesterol in the phospholipid membrane affect the spectroscopic properties of lipophilic dye nile red

Science.gov (United States)

Halder, Animesh; Saha, Baishakhi; Maity, Pabitra; Kumar, Gopinatha Suresh; Sinha, Deepak Kumar; Karmakar, Sanat

2018-02-01

We have studied the effect of composition and the phase state of phospholipid membranes on the emission spectrum, anisotropy and lifetime of a lipophilic fluorescence probe nile red. Fluorescence spectrum of nile red in membranes containing cholesterol has also been investigated in order to get insights into the influence of cholesterol on the phospholipid membranes. Maximum emission wavelength (λem) of nile red in the fluid phase of saturated and unsaturated phospholipids was found to differ by 10 nm. The λem was also found to be independent of chain length and charge of the membrane. However, the λem is strongly dependent on the temperature in the gel phase. The λem and rotational diffusion rate decrease, whereas the anisotropy and lifetime increase markedly with increasing cholesterol concentration for saturated phosoholipids, such as, dimyristoyl phosphatidylcholine (DMPC) in the liquid ordered phase. However, these spectroscopic properties do not alter significantly in case of unsaturated phospholipids, such as, dioleoyl phosphatidylcholine (DOPC) in liquid disordered phase. Interestingly, red edge excitation shift (REES) in the presence of lipid-cholesterol membranes is the direct consequences of change in rotational diffusion due to motional restriction of lipids in the presence of cholesterol. This study provides correlations between the membrane compositions and fluorescence spectral features which can be utilized in a wide range of biophysical fields as well the cell biology.

Biophysical studies of cholesterol in unsaturated phospholipid model membranes

Science.gov (United States)

Williams, Justin Adam

Cellular membranes contain a staggering diversity of lipids. The lipids are heterogeneously distributed to create regions, or domains, whose physical properties differ from the bulk membrane and play an essential role in modulating the function of resident proteins. Many basic questions pertaining to the formation of these lateral assemblies remain. This research employs model membranes of well-defined composition to focus on the potential role of polyunsaturated fatty acids (PUFAs) and their interaction with cholesterol (chol) in restructuring the membrane environment. Omega-3 (n-3) PUFAs are the main bioactive components of fish oil, whose consumption alleviates a variety of health problems by a molecular mechanism that is unclear. We hypothesize that the incorporation of PUFAs into membrane lipids and the effect they have on molecular organization may be, in part, responsible. Chol is a major constituent in the plasma membrane of mammals. It determines the arrangement and collective properties of neighboring lipids, driving the formation of domains via differential affinity for different lipids. The molecular organization of 1-[2H31]palmitoyl-2-eicosapentaenoylphosphatidylcholine (PEPC-d31) and 1-[2H31]palmitoyl-2-docosahexaenoylphosphatidylcholine (PDPC-d31) in membranes with sphingomyelin (SM) and chol (1:1:1 mol) was compared by solid-state 2H NMR spectroscopy. Eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids are the two major n-3 PUFAs found in fish oil, while PEPC-d31 and PDPC-d31 are phospholipids containing the respective PUFAs at the sn-2 position and a perdeuterated palmitic acid at the sn-1 position. Analysis of spectra recorded as a function of temperature indicates that in both cases, formation of PUFA-rich (less ordered) and SM-rich (more ordered) domains occurred. A surprisingly substantial proportion of PUFA was found to infiltrate the more ordered domain. There was almost twice as much DHA (65%) as EPA (30%). The implication is that n-3

Phospholipid composition and a polybasic motif determine D6 PROTEIN KINASE polar association with the plasma membrane and tropic responses.

Science.gov (United States)

Barbosa, Inês C R; Shikata, Hiromasa; Zourelidou, Melina; Heilmann, Mareike; Heilmann, Ingo; Schwechheimer, Claus

2016-12-15

Polar transport of the phytohormone auxin through PIN-FORMED (PIN) auxin efflux carriers is essential for the spatiotemporal control of plant development. The Arabidopsis thaliana serine/threonine kinase D6 PROTEIN KINASE (D6PK) is polarly localized at the plasma membrane of many cells where it colocalizes with PINs and activates PIN-mediated auxin efflux. Here, we show that the association of D6PK with the basal plasma membrane and PINs is dependent on the phospholipid composition of the plasma membrane as well as on the phosphatidylinositol phosphate 5-kinases PIP5K1 and PIP5K2 in epidermis cells of the primary root. We further show that D6PK directly binds polyacidic phospholipids through a polybasic lysine-rich motif in the middle domain of the kinase. The lysine-rich motif is required for proper PIN3 phosphorylation and for auxin transport-dependent tropic growth. Polybasic motifs are also present at a conserved position in other D6PK-related kinases and required for membrane and phospholipid binding. Thus, phospholipid-dependent recruitment to membranes through polybasic motifs might not only be required for D6PK-mediated auxin transport but also other processes regulated by these, as yet, functionally uncharacterized kinases. © 2016. Published by The Company of Biologists Ltd.

Anhydrous ethanol: A renewable source of energy

Energy Technology Data Exchange (ETDEWEB)

Kumar, Santosh; Singh, Neetu; Prasad, Ram [Department of Chemical Engineering, H. B. Technological Institute, Kanpur 208002 (India)

2010-09-15

Anhydrous ethanol is one of the biofuels produced today and it is a subset of renewable energy. It is considered to be an excellent alternative clean-burning fuel to gasoline. Anhydrous ethanol is commercially produced by either catalytic hydration of ethylene or fermentation of biomass. Any biological material that has sugar, starch or cellulose can be used as biomass for producing anhydrous ethanol. Since ethanol-water solution forms a minimum-boiling azeotrope of composition of 89.4 mol% ethanol and 10.6 mol% water at 78.2 C and standard atmospheric pressure, the dilute ethanol-water solutions produced by fermentation process can be continuously rectified to give at best solutions containing 89.4 mol% ethanol at standard atmospheric pressure. Therefore, special process for removal of the remaining water is required for manufacture of anhydrous ethanol. Various processes for producing anhydrous ethanol have been used/suggested. These include: (i) chemical dehydration process, (ii) dehydration by vacuum distillation process, (iii) azeotropic distillation process, (iv) extractive distillation processes, (v) membrane processes, (vi) adsorption processes and (vii) diffusion distillation process. These processes of manufacturing anhydrous ethanol have been improved continuously due to the increasingly strict requirements for quantity and quality of this product. The literature available on these processes is reviewed. These processes are also compared on the basis of energy requirements. (author)

Role of phospholipids in destabilization of lysosomal membranes in chronic alcohol poisoning

International Nuclear Information System (INIS)

Tadevosyan, Y.V.; Batikyan, T.B.; Gevorkyan, G.A.; Karagezyan, K.G.

1986-01-01

The aim of this investigation was to study changes in the phospholipids (PL) spectrum and possible activity of membrane-bound phospholipase A 2 in lysosomal membranes from albino rat liver under conditions of the normally metabolizing tissue and during long-term alcohol poisoning. Changes in stability of the lysosomal membranes were determined by measuring nonsedimented acid phosphatase (AP) activity. The substance 1-acyl-2-(1- 14 C)-oleoyl-phosphatidyl-choline ( 14 C-PCh) was synthesized by an enzymic method. Phospholipase A 2 activity was determined in an incubation medium of Tris-Maleate buffer containing 20 nanomoles ( 14 C)-PCH, 8 mM CaC1 2 , and about 100 micrograms protein

Relation between various phospholipase actions on human red cell membranes and the interfacial phospholipid pressure in monolayers

NARCIS (Netherlands)

Demel, R.A.; Geurts van Kessel, W.S.M.; Zwaal, R.F.A.; Roelofsen, B.; Deenen, L.L.M. van

1975-01-01

The action of purified phospholipases on monomolecular films of various interfacial pressures is compared with the action on erythrocyte membranes. The phospholipases which cannot hydrolyse phospholipids of the intact erythrocyte membrane, phospholipase C from Bacillus cereus, phospholipase A2 from

Membrane phospholipid composition may contribute to exceptional longevity of the naked mole-rat (Heterocephalus glaber): a comparative study using shotgun lipidomics.

Science.gov (United States)

Mitchell, Todd W; Buffenstein, Rochelle; Hulbert, A J

2007-11-01

Phospholipids containing highly polyunsaturated fatty acids are particularly prone to peroxidation and membrane composition may therefore influence longevity. Phospholipid molecules, in particular those containing docosahexaenoic acid (DHA), from the skeletal muscle, heart, liver and liver mitochondria were identified and quantified using mass-spectrometry shotgun lipidomics in two similar-sized rodents that show an approximately 9-fold difference in maximum lifespan. The naked mole rat is the longest-living rodent known with a maximum lifespan of >28 years. Total phospholipid distribution is similar in tissues of both species; DHA is only found in phosphatidylcholines (PC), phosphatidylethanolamines (PE) and phosphatidylserines (PS), and DHA is relatively more concentrated in PE than PC. Naked mole-rats have fewer molecular species of both PC and PE than do mice. DHA-containing phospholipids represent 27-57% of all phospholipids in mice but only 2-6% in naked mole-rats. Furthermore, while mice have small amounts of di-polyunsaturated PC and PE, these are lacking in naked mole-rats. Vinyl ether-linked phospholipids (plasmalogens) are higher in naked mole-rat tissues than in mice. The lower level of DHA-containing phospholipids suggests a lower susceptibility to peroxidative damage in membranes of naked mole-rats compared to mice. Whereas the high level of plasmalogens might enhance membrane antioxidant protection in naked mole-rats compared to mice. Both characteristics possibly contribute to the exceptional longevity of naked mole-rats and may indicate a special role for peroxisomes in this extended longevity.

(CryoTransmission Electron Microscopy of Phospholipid Model Membranes Interacting with Amphiphilic and Polyphilic Molecules

Directory of Open Access Journals (Sweden)

Annette Meister

2017-10-01

Full Text Available Lipid membranes can incorporate amphiphilic or polyphilic molecules leading to specific functionalities and to adaptable properties of the lipid bilayer host. The insertion of guest molecules into membranes frequently induces changes in the shape of the lipid matrix that can be visualized by transmission electron microscopy (TEM techniques. Here, we review the use of stained and vitrified specimens in (cryoTEM to characterize the morphology of amphiphilic and polyphilic molecules upon insertion into phospholipid model membranes. Special emphasis is placed on the impact of novel synthetic amphiphilic and polyphilic bolalipids and polymers on membrane integrity and shape stability.

Improved mitochondrial function with diet-induced increase in either docosahexaenoic acid or arachidonic acid in membrane phospholipids.

Directory of Open Access Journals (Sweden)

Ramzi J Khairallah

Full Text Available Mitochondria can depolarize and trigger cell death through the opening of the mitochondrial permeability transition pore (MPTP. We recently showed that an increase in the long chain n3 polyunsaturated fatty acids (PUFA docosahexaenoic acid (DHA; 22:6n3 and depletion of the n6 PUFA arachidonic acid (ARA; 20:4n6 in mitochondrial membranes is associated with a greater Ca(2+ load required to induce MPTP opening. Here we manipulated mitochondrial phospholipid composition by supplementing the diet with DHA, ARA or combined DHA+ARA in rats for 10 weeks. There were no effects on cardiac function, or respiration of isolated mitochondria. Analysis of mitochondrial phospholipids showed DHA supplementation increased DHA and displaced ARA in mitochondrial membranes, while supplementation with ARA or DHA+ARA increased ARA and depleted linoleic acid (18:2n6. Phospholipid analysis revealed a similar pattern, particularly in cardiolipin. Tetralinoleoyl cardiolipin was depleted by 80% with ARA or DHA+ARA supplementation, with linoleic acid side chains replaced by ARA. Both the DHA and ARA groups had delayed Ca(2+-induced MPTP opening, but the DHA+ARA group was similar to the control diet. In conclusion, alterations in mitochondria membrane phospholipid fatty acid composition caused by dietary DHA or ARA was associated with a greater cumulative Ca(2+ load required to induced MPTP opening. Further, high levels of tetralinoleoyl cardiolipin were not essential for normal mitochondrial function if replaced with very-long chain n3 or n6 PUFAs.

Ultrafiltration-based degumming of crude rice bran oil using a polymer membrane

Energy Technology Data Exchange (ETDEWEB)

Sehn, G.A.R.; GonCalves, L.A.G.; Ming, C.C.

2016-07-01

Membrane technology has been gaining momentum in industrial processes, especially in food technology. It is believed to simplify processes, reduce energy consumption, and eliminate pollutants. The objective was to study the performance of polyvinylidene fluoride (PVDF) and polyethersulfone (PES) polymeric membranes in the degumming of the miscella of crude rice bran oil by using a bench-scale tangential filtration module. In addition, oil miscella filtration techniques using hexane and anhydrous ethyl alcohol solvents were compared. All membranes showed the retention of phospholipids and high flow rates. However, the best performance was observed using the 50-kDa PVDF membrane in miscella hexane solvent, with a 95.5% retention of the phosphorus concentration (by a factor of 1.4), resulting in a permeate with 29 mg·kg−1 of phosphorus and an average flow rate of 48.1 L·m−2·h−1. This technology can be used as a low-pollution, economical alternative for the de-gumming of crude rice bran oil, being effective in the removal of hydratable and non-hydratable phospholipids, resulting in oils with a low phosphorus content. (Author)

Herpes simplex virus 1 induces de novo phospholipid synthesis

Energy Technology Data Exchange (ETDEWEB)

Sutter, Esther [Electron Microscopy, Institute of Veterinary Anatomy, University of Zuerich (Switzerland); Oliveira, Anna Paula de; Tobler, Kurt [Electron microscopy, Institute of Virology, University of Zuerich (Switzerland); Schraner, Elisabeth M. [Electron Microscopy, Institute of Veterinary Anatomy, University of Zuerich (Switzerland); Sonda, Sabrina [Institute of Parasitology, University of Zuerich (Switzerland); Kaech, Andres [Center for Microscopy and Image Analysis, University of Zuerich (Switzerland); Lucas, Miriam S. [Electron Microscopy ETH Zuerich (EMEZ), Swiss Federal Institute of Technology, Zuerich (Switzerland); Ackermann, Mathias [Electron microscopy, Institute of Virology, University of Zuerich (Switzerland); Wild, Peter, E-mail: pewild@access.uzh.ch [Electron Microscopy, Institute of Veterinary Anatomy, University of Zuerich (Switzerland)

2012-08-01

Herpes simplex virus type 1 capsids bud at nuclear membranes and Golgi membranes acquiring an envelope composed of phospholipids. Hence, we measured incorporation of phospholipid precursors into these membranes, and quantified changes in size of cellular compartments by morphometric analysis. Incorporation of [{sup 3}H]-choline into both nuclear and cytoplasmic membranes was significantly enhanced upon infection. [{sup 3}H]-choline was also part of isolated virions even grown in the presence of brefeldin A. Nuclei expanded early in infection. The Golgi complex and vacuoles increased substantially whereas the endoplasmic reticulum enlarged only temporarily. The data suggest that HSV-1 stimulates phospholipid synthesis, and that de novo synthesized phospholipids are inserted into nuclear and cytoplasmic membranes to i) maintain membrane integrity in the course of nuclear and cellular expansion, ii) to supply membrane constituents for envelopment of capsids by budding at nuclear membranes and Golgi membranes, and iii) to provide membranes for formation of transport vacuoles.

Effect of heavy water on phospholipid membranes: experimental confirmation of molecular dynamics simulations

Czech Academy of Sciences Publication Activity Database

Beranová, Lenka; Humpolíčková, Jana; Sýkora, Jan; Benda, Aleš; Cwiklik, Lukasz; Jurkiewicz, Piotr; Gröbner, G.; Hof, Martin

Roč. 14, č. 42 ( 2012 ), s. 14516-14522 ISSN 1463-9076 R&D Projects: GA AV ČR GEMEM/09/E006; GA ČR GBP208/12/G016 Institutional support: RVO:61388955 Keywords : phospholipid membranes * biophysics * physical chemistry Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.829, year: 2012

Parallel artificial liquid membrane extraction as an efficient tool for removal of phospholipids from human plasma.

Science.gov (United States)

Ask, Kristine Skoglund; Bardakci, Turgay; Parmer, Marthe Petrine; Halvorsen, Trine Grønhaug; Øiestad, Elisabeth Leere; Pedersen-Bjergaard, Stig; Gjelstad, Astrid

2016-09-10

Generic Parallel Artificial Liquid Membrane Extraction (PALME) methods for non-polar basic and non-polar acidic drugs from human plasma were investigated with respect to phospholipid removal. In both cases, extractions in 96-well format were performed from plasma (125μL), through 4μL organic solvent used as supported liquid membranes (SLMs), and into 50μL aqueous acceptor solutions. The acceptor solutions were subsequently analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) using in-source fragmentation and monitoring the m/z 184→184 transition for investigation of phosphatidylcholines (PC), sphingomyelins (SM), and lysophosphatidylcholines (Lyso-PC). In both generic methods, no phospholipids were detected in the acceptor solutions. Thus, PALME appeared to be highly efficient for phospholipid removal. To further support this, qualitative (post-column infusion) and quantitative matrix effects were investigated with fluoxetine, fluvoxamine, and quetiapine as model analytes. No signs of matrix effects were observed. Finally, PALME was evaluated for the aforementioned drug substances, and data were in accordance with European Medicines Agency (EMA) guidelines. Copyright © 2016 Elsevier B.V. All rights reserved.

A new anhydrous proton conductor based on polybenzimidazole and tridecyl phosphate

International Nuclear Information System (INIS)

Jiang Fengjing; Pu Hongting; Meyer, Wolfgang H.; Guan Yisi; Wan Decheng

2008-01-01

Most of the anhydrous proton conducting membranes are based on inorganic or partially inorganic materials, like SrCeO 3 membranes or polybenzimidazole (PBI)/H 3 PO 4 composite membranes. In present work, a new kind of anhydrous proton conducting membrane based on fully organic components of PBI and tridecyl phosphate (TP) was prepared. The interaction between PBI and TP is discussed. The temperature dependence of the proton conductivity of the composite membranes can be modeled by an Arrhenius relation. Thermogravimetric analysis (TGA) illustrates that these composite membranes are chemically stable up to 145 deg. C. The weight loss appearing at 145 deg. C is attributed to the selfcondensation of phosphate, which results in the proton conductivity drop of the membranes occurring at the same temperature. The DC conductivity of the composite membranes can reach ∼10 -4 S/cm for PBI/1.8TP at 140 deg. C and increases with increasing TP content. The proton conductivity of PBI/TP and PBI/H 3 PO 4 composite membranes is compared. The former have higher proton conductivity, however, the proton conductivity of the PBI/H 3 PO 4 membranes increases with temperature more significantly. Compared with PBI/H 3 PO 4 membranes, the migration stability of TP in PBI/TP membranes is improved significantly

Dicarboxylic phospholipids and irradiated biomembranes

International Nuclear Information System (INIS)

Dousset, Nicole.

1977-01-01

It was decided to study the effects of ionizing radiations on biomembranes, with special reference to erythrocytes and liver microsomes representing two kinds of membrane very common in nature. Diacid phospholipids were observed at these membranes and the results are reported in part one of this work. It appeared essential to examine as far as possible the metabolism, in vitro and in animals, of these diacids and to find out whether certain harmful effects of radiations on the proteins (membrane permeability changes and enzyme inactivation) could be due to the action of these newly formed compounds. The study of acid compounds formed under irradiation was limited to nonanal-9-oic acid and azelaic acid. Part two deals with the incorporation of acid and diacid compounds into lipids and the effects of diacid phospholipids on the membrane permeability. A chapter is devoted to the changes in certain enzyme activities brought about by diacid phospholipids [fr

Perimicrovillar membrane assembly: the fate of phospholipids synthesised by the midgut of Rhodnius prolixus

Directory of Open Access Journals (Sweden)

Paula Rego Bittencourt-Cunha

2013-06-01

Full Text Available In this study, we describe the fate of fatty acids that are incorporated from the lumen by the posterior midgut epithelium of Rhodnius prolixus and the biosynthesis of lipids. We also demonstrate that neutral lipids (NL are transferred to the haemolymphatic lipophorin (Lp and that phospholipids remain in the tissue in which they are organised into perimicrovillar membranes (PMMs. 3H-palmitic acid added at the luminal side of isolated midguts of R. prolixus females was readily absorbed and was used to synthesise phospholipids (80% and NL (20%. The highest incorporation of 3H-palmitic acid was on the first day after a blood meal. The amounts of diacylglycerol (DG and triacylglycerol synthesised by the tissue decreased in the presence of Lp in the incubation medium. The metabolic fates of 3H-lipids synthesised by the posterior midgut were followed and it was observed that DG was the major lipid released to Lp particles. However, the majority of phospholipids were not transferred to Lp, but remained in the tissue. The phospholipids that were synthesised and accumulated in the posterior midgut were found to be associated with Rhodnius luminal contents as structural components of PMMs.

Confocal Raman Microscopy for in Situ Measurement of Phospholipid-Water Partitioning into Model Phospholipid Bilayers within Individual Chromatographic Particles.

Science.gov (United States)

Kitt, Jay P; Bryce, David A; Minteer, Shelley D; Harris, Joel M

2018-06-05

The phospholipid-water partition coefficient is a commonly measured parameter that correlates with drug efficacy, small-molecule toxicity, and accumulation of molecules in biological systems in the environment. Despite the utility of this parameter, methods for measuring phospholipid-water partition coefficients are limited. This is due to the difficulty of making quantitative measurements in vesicle membranes or supported phospholipid bilayers, both of which are small-volume phases that challenge the sensitivity of many analytical techniques. In this work, we employ in situ confocal Raman microscopy to probe the partitioning of a model membrane-active compound, 2-(4-isobutylphenyl) propionic acid or ibuprofen, into both hybrid- and supported-phospholipid bilayers deposited on the pore walls of individual chromatographic particles. The large surface-area-to-volume ratio of chromatographic silica allows interrogation of a significant lipid bilayer area within a very small volume. The local phospholipid concentration within a confocal probe volume inside the particle can be as high as 0.5 M, which overcomes the sensitivity limitations of making measurements in the limited membrane areas of single vesicles or planar supported bilayers. Quantitative determination of ibuprofen partitioning is achieved by using the phospholipid acyl-chains of the within-particle bilayer as an internal standard. This approach is tested for measurements of pH-dependent partitioning of ibuprofen into both hybrid-lipid and supported-lipid bilayers within silica particles, and the results are compared with octanol-water partitioning and with partitioning into individual optically trapped phospholipid vesicle membranes. Additionally, the impact of ibuprofen partitioning on bilayer structure is evaluated for both within-particle model membranes and compared with the structural impacts of partitioning into vesicle lipid bilayers.

Lipophilic organic pollutants induce changes in phospholipid and membrane protein composition leading to Vero cell morphological change.

Science.gov (United States)

Liao, Ting T; Wang, Lei; Jia, Ru W; Fu, Xiao H; Chua, Hong

2014-01-01

Membrane damage related to morphological change in Vero cells is a sensitive index of the composite biotoxicity of trace lipophilic chemicals. However, judging whether the morphological change in Vero cells happens and its ratio are difficult because it is not a quantitative characteristic. To find biomarkers of cell morphological change for quantitatively representing the ratio of morphological changed cell, the mechanism of cell membrane damage driven by typical lipophilic chemicals, such as trichlorophenol (TCP) and perfluorooctanesulphonate (PFOS), was explored. The ratio of morphologically changed cells generally increased with increased TCP or PFOS concentrations, and the level of four major components of phospholipids varied with concentrations of TCP or PFOS, but only the ratio of phosphatidylcholine (PC)/phosphatidylethanolamine (PE) decreased regularly as TCP or PFOS concentrations increased. Analysis of membrane proteins showed that the level of vimentin in normal cell membranes is high, while it decreases or vanishes after TCP exposure. These variations in phospholipid and membrane protein components may result in membrane leakage and variation in rigid structure, which leads to changes in cell morphology. Therefore, the ratio of PC/PE and amount of vimentin may be potential biomarkers for representing the ratio of morphological changed Vero cell introduced by trace lipophilic compounds, thus their composite bio-toxicity.

Morphological and physical analysis of natural phospholipids-based biomembranes.

Directory of Open Access Journals (Sweden)

Adrien Jacquot

Full Text Available BACKGROUND: Liposomes are currently an important part of biological, pharmaceutical, medical and nutritional research, as they are considered to be among the most effective carriers for the introduction of various types of bioactive agents into target cells. SCOPE OF REVIEW: In this work, we study the lipid organization and mechanical properties of biomembranes made of marine and plant phospholipids. Membranes based on phospholipids extracted from rapeseed and salmon are studied in the form of liposome and as supported lipid bilayer. Dioleylphosphatidylcholine (DOPC and dipalmitoylphosphatidylcholine (DPPC are used as references to determine the lipid organization of marine and plant phospholipid based membranes. Atomic force microscopy (AFM imaging and force spectroscopy measurements are performed to investigate the membranes' topography at the micrometer scale and to determine their mechanical properties. MAJOR CONCLUSIONS: The mechanical properties of the membranes are correlated to the fatty acid composition, the morphology, the electrophoretic mobility and the membrane fluidity. Thus, soft and homogeneous mechanical properties are evidenced for salmon phospholipids membrane containing various polyunsaturated fatty acids. Besides, phase segregation in rapeseed membrane and more important mechanical properties were emphasized for this type of membranes by contrast to the marine phospholipids based membranes. GENERAL SIGNIFICANCE: This paper provides new information on the nanomechanical and morphological properties of membrane in form of liposome by AFM. The originality of this work is to characterize the physico-chemical properties of the nanoliposome from the natural sources containing various fatty acids and polar head.

Physiological levels of diacylglycerols in phospholipid membranes induce membrane fusion and stabilize inverted phases

International Nuclear Information System (INIS)

Siegel, D.P.; Banschbach, J.; Alford, D.; Ellens, H.; Lis, L.J.; Quinn, P.J.; Yeagle, P.L.; Bentz, J.

1989-01-01

In a previous paper, it was shown that liposome fusion rates are substantially enhanced under the same conditions which induce isotropic 31 P NMR resonances in multilamellar dispersions of the same lipid. Both of these phenomena occur within the same temperature interval, ΔT I , below the L α /H II phase transition temperature, T H . T H and ΔT I can be extremely sensitive to the lipid composition. The present work shows that 2 mol % of diacylglycerols like those produced by the phosphatidylinositol cycle in vivo can lower T H , ΔT I , and the temperature for fast membrane fusion by 15-20 degree C. N-Monomethylated dioleoylphosphatidylethanolamine is used as a model system. These results show that physiological levels of diacylglycerols can substantially increase the susceptibility of phospholipid membranes to fusion. This suggests that, in addition to their role in protein kinase C activation, diacylglycerols could play a more direct role in the fusion event during stimulus-exocytosis coupling in vivo

Effects of Chain Length and Saturability of Fatty Acids on Phospholipids and Proteins in Plasma Membranes of Bovine Mammary Gland.

Science.gov (United States)

Yan, Qiongxian; Tang, Shaoxun; Han, Xuefeng; Bamikole, Musibau Adungbe; Zhou, Chuanshe; Kang, Jinhe; Wang, Min; Tan, Zhiliang

2016-12-01

Free fatty acids (FFAs) in plasma are essential substrates for de novo synthesis of milk fat, or directly import into mammary cells. The physico-chemical properties of mammary cells membrane composition affected by FFAs with different chain lengths and saturability are unclear yet. Employing GC, FTIR and fluorescence spectroscopy, the adsorption capacity, phospholipids content, membrane proteins conformation, lipid peroxidation product, and free sulfhydryl of plasma membranes (PMs) interacted with different FFAs were determined. The mammary cells PMs at 38 and 39.5 °C showed different adsorption capacities: acetic acid (Ac) > stearic acid (SA) > β-hydroxybutyric acid (BHBA) > trans10, cis12 CLA. In the FTIR spectrum, the major adsorption peaks appeared at 2920 and 2850 cm -1 for phospholipids, and at 1628 and 1560 cm -1 for membrane proteins. The intensities of PMs-FFAs complexes were varied with the FFAs species and their initial concentrations. The β-sheet and turn structures of membrane proteins were transferred into random coil and α-helix after BHBA, SA and trans10, cis12 CLA treatments compared with Ac treatment. The quenching effects on the fluorescence of endogenous membrane protein, 1, 8-ANS, NBD-PE, and DHPE entrapped in PMs by LCFA were different from those of short chain FFAs. These results indicate that the adsorption of FFAs could change membrane protein conformation and polarity of head group in phospholipids. This variation of the mammary cells PMs was regulated by carbon chain length and saturability of FFAs.

Proceedings of the users meeting on structure and phase transition of phospholipid membrane

International Nuclear Information System (INIS)

Hatta, Ichiro; Amemiya, Yoshiyuki

1994-06-01

On the occasion that the persons of three groups that have carried out the research on the structure and the phase transition of phospholipid membranes have carried out the experiment successively, the users meeting was held on November 1, 1993 at National Laboratory for High Energy Physics. Lectures were given on the L βI structure of DPPC/alcohol system, the self gathering and intermolecular cooperation phenomenon of glycero phospholipid, the phase transition of DEPE/water system, the structure of DMPA/polylysine, the development of X-ray television, the ripple structure of DMPC/cholesterol system and the simultaneous measurement of X-ray diffraction/DSC. To have the chance like this is very meaningful because sufficient discussion can be done among usually busy researchers at the synchrotron radiation experiment facility. (K.I.)

Proceedings of the users meeting on structure and phase transition of phospholipid membrane

Energy Technology Data Exchange (ETDEWEB)

Hatta, Ichiro [Nagoya Univ. (Japan). School of Engineering; Amemiya, Yoshiyuki [eds.

1994-06-01

On the occasion that the persons of three groups that have carried out the research on the structure and the phase transition of phospholipid membranes have carried out the experiment successively, the users meeting was held on November 1, 1993 at National Laboratory for High Energy Physics. Lectures were given on the L{sub {beta}I} structure of DPPC/alcohol system, the self gathering and intermolecular cooperation phenomenon of glycero phospholipid, the phase transition of DEPE/water system, the structure of DMPA/polylysine, the development of X-ray television, the ripple structure of DMPC/cholesterol system and the simultaneous measurement of X-ray diffraction/DSC. To have the chance like this is very meaningful because sufficient discussion can be done among usually busy researchers at the synchrotron radiation experiment facility. (K.I.).

Curcumin liposomes prepared with milk fat globule membrane phospholipids and soybean lecithin.

Science.gov (United States)

Jin, Hong-Hao; Lu, Qun; Jiang, Jian-Guo

2016-03-01

Using thin film ultrasonic dispersion method, the curcumin liposomes were prepared with milk fat globule membrane (MFGM) phospholipids and soybean lecithins, respectively, to compare the characteristics and stability of the 2 curcumin liposomes. The processing parameters of curcumin liposomes were investigated to evaluate their effects on the encapsulation efficiency. Curcumin liposomes were characterized in terms of size distribution, ζ-potential, and in vitro release behavior, and then their storage stability under various conditions was evaluated. The curcumin liposomes prepared with MFGM phospholipids had an encapsulation efficiency of about 74%, an average particle size of 212.3 nm, and a ζ-potential of -48.60 mV. The MFGM liposomes showed higher encapsulation efficiency, smaller particle size, higher absolute value of ζ-potential, and slower in vitro release than soybean liposomes. The retention rate of liposomal curcumin was significantly higher than that of free curcumin. The stability of the 2 liposomes under different pH was almost the same, but MFGM liposomes displayed a slightly higher stability than soybean liposomes under the conditions of Fe(3+), light, temperature, oxygen, and relative humidity. In conclusion, MFGM phospholipids have potential advantages in the manufacture of curcumin liposomes used in food systems. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Control of phospholipid flip-flop by transmembrane peptides

International Nuclear Information System (INIS)

Kaihara, Masanori; Nakao, Hiroyuki; Yokoyama, Hirokazu; Endo, Hitoshi; Ishihama, Yasushi; Handa, Tetsurou; Nakano, Minoru

2013-01-01

Highlights: ► Phospholipid flip-flop in transmembrane peptide-containing vesicles was investigated. ► Peptides that contained polar residues in the center of the transmembrane region promoted phospholipid flip-flop. ► A bioinformatics approach revealed the presence of polar residues in the transmembrane region of ER membrane proteins. ► Polar residues in ER membrane proteins possibly provide flippase-like activity. - Abstract: We designed three types of transmembrane model peptides whose sequence originates from a frequently used model peptide KALP23, and we investigated their effects on phospholipid flip-flop. Time-resolved small-angle neutron scattering and a dithionite fluorescent quenching assay demonstrated that TMP-L, which has a fully hydrophobic transmembrane region, did not enhance phospholipid flip-flop, whereas TMP-K and TMP-E, which have Lys and Glu, respectively, in the center of their transmembrane regions, enhanced phospholipid flip-flop. Introduction of polar residues in the membrane-spanning helices is considered to produce a locally polar region and enable the lipid head group to interact with the polar side-chain inside the bilayers, thereby reducing the activation energy for the flip-flop. A bioinformatics approach revealed that acidic and basic residues account for 4.5% of the central region of the transmembrane domain in human ER membrane proteins. Therefore, polar residues in ER membrane proteins are considered to provide flippase-like activity

Parallel artificial liquid membrane extraction as an efficient tool for removal of phospholipids from human plasma

DEFF Research Database (Denmark)

Ask, Kristine Skoglund; Bardakci, Turgay; Parmer, Marthe Petrine

2016-01-01

Generic Parallel Artificial Liquid Membrane Extraction (PALME) methods for non-polar basic and non-polar acidic drugs from human plasma were investigated with respect to phospholipid removal. In both cases, extractions in 96-well format were performed from plasma (125μL), through 4μL organic...

Seasonal changes in minor membrane phospholipid classes, sterols and tocopherols in overwintering insect, Pyrrhocoris apterus

Czech Academy of Sciences Publication Activity Database

Košťál, Vladimír; Urban, T.; Řimnáčová, Lucie; Berková, Petra; Šimek, Petr

2013-01-01

Roč. 59, č. 9 (2013), s. 934-941 ISSN 0022-1910 R&D Projects: GA MŠk LH12103; GA MZd(CZ) NT11513 Institutional support: RVO:60077344 Keywords : seasonal membrane restructuring * phospholipids * lysophospholipids Subject RIV: ED - Physiology Impact factor: 2.500, year: 2013 http://www.sciencedirect.com/science/article/pii/S0022191013001406#

Temperature-induced changes in lecithin model membranes detected by novel covalent spin-labelled phospholipids.

Science.gov (United States)

Stuhne-Sekalec, L; Stanacev, N Z

1977-02-01

Several spin-labelled phospholipids carrying covalently bound 5-doxylstearic acid (2-(3-carboxydecyl)-2-hexyl-4,4-dimethyl-3-oxazolidinoxyl) were intercalated in liposomes of saturated and unsaturated lecithins. Temperature-induced changes of these liposomes, detected by the spin-labelled phospholipids, were found to be in agreement with the previously described transitions of hydrocarbon chains of host lecithins detected by different probes and different techniques, establishing that spin-labelled phosopholipids are sensitive probes for the detection of temperature-induced changes in lecithin model membranes. In addition to the detection of already-known transitions in lecithin liposomes, the coexistence of two distinctly different enviroments was observed above the characteristic transition temperature. This phenomenon was tentatively attributed to the influence of the lecithin polar group on the fluidity of fatty acyl chains near the polar group. Combined with other results from the literature, the coexistence of two environments could be associated with the coexistence of two conformational isomers of lecithin, differing in the orientation of the polar head group with respect to the plane of bilayer. These findings have been discussed in view of the present state of knowledge regarding temperature-induced changes in model membranes.

Polyhydroxyalkanoate (PHA) Granules Have no Phospholipids

Science.gov (United States)

Bresan, Stephanie; Sznajder, Anna; Hauf, Waldemar; Forchhammer, Karl; Pfeiffer, Daniel; Jendrossek, Dieter

2016-01-01

Polyhydroxybutyrate (PHB) granules, also designated as carbonosomes, are supra-molecular complexes in prokaryotes consisting of a PHB polymer core and a surface layer of structural and functional proteins. The presence of suspected phospholipids in the surface layer is based on in vitro data of isolated PHB granules and is often shown in cartoons of the PHB granule structure in reviews on PHB metabolism. However, the in vivo presence of a phospholipid layer has never been demonstrated. We addressed this topic by the expression of fusion proteins of DsRed2EC and other fluorescent proteins with the phospholipid-binding domain (LactC2) of lactadherin in three model organisms. The fusion proteins specifically localized at the cell membrane of Ralstonia eutropha but did not co-localize with PHB granules. The same result was obtained for Pseudomonas putida, a species that accumulates another type of polyhydroxyalkanoate (PHA) granules related to PHB. Notably, DsRed2EC-LactC2 expressed in Magnetospirillum gryphiswaldense was detected at the position of membrane-enclosed magnetosome chains and at the cytoplasmic membrane but not at PHB granules. In conclusion, the carbonosomes of representatives of α-proteobacteria, β-proteobacteria and γ-proteobacteria have no phospholipids in vivo and we postulate that the PHB/PHA granule surface layers in natural producers generally are free of phospholipids and consist of proteins only. PMID:27222167

Model mass spectrometric study of competitive interactions of antimicrobial bisquaternary ammonium drugs and aspirin with membrane phospholipids

Directory of Open Access Journals (Sweden)

Vekey K.

2013-03-01

Full Text Available The aim of the study is to reveal molecular mechanisms of possible activity modulation of antimicrobial bis-quaternary ammonium compounds (BQAC and aspirin (ASP through noncovalent competitive complexation under their combined introduction into the model systems with membrane phospholipids. Methods. Binary and triple systems containing either decamethoxinum or ethonium, or thionium and aspirin, as well as dipalmitoyl-phosphatidylcholine (DPPC have been investigated by electrospray ionization mass spectrometry. Results. Basing on the analysis of associates recorded in the mass spectra, the types of nonocovalent complexes formed in the systems studied were determined and the supposed role of the complexation in the BQAC and ASP activity modulation was discussed. The formation of associates of BQAC dications with ASP anion is considered as one of the possible ways of deactivation of ionic forms of the medications. The formation of stable complexes of BQAC with DPPC and ASP with DPPC in binary systems as well as the complexes distribution in triple-components systems BQAC:ASP:DPPC point to the existence of competition between drugs of these two types for the binding to DPPC. Conclusions. The results obtained point to the competitive complexation in the model molecular systems containing the BQAC, aspirin and membrane phospholipids. The observed phenomenon testifies to the possibility of modulating the activity of bisquaternary antimicrobial agents and aspirin under their combined usage, due to the competition between the drugs for binding to the target membrane phospholipid molecules and also due to the formation of stable noncovalent complexes between BQAC and ASP.

Molecular phospholipid films on solid supports

DEFF Research Database (Denmark)

Czolkos, Ilja; Jesorka, Aldo; Orwar, Owe

2011-01-01

Phospholipid membranes are versatile structures for mimicking biological surfaces. Bilayer and monolayer membranes can be formed on solid supports, leading to enhanced stability and accessibility of the biomimetic molecular film. This has facilitated functional studies of membrane proteins and ai...

Protein kinase C interaction with calcium: a phospholipid-dependent process.

LENUS (Irish Health Repository)

Bazzi, M D

1990-08-21

The calcium-binding properties of calcium- and phospholipid-dependent protein kinase C (PKC) were investigated by equilibrium dialysis in the presence and the absence of phospholipids. Calcium binding to PKC displayed striking and unexpected behavior; the free proteins bound virtually no calcium at intracellular calcium concentrations and bound limited calcium (about 1 mol\\/mol of PKC) at 200 microM calcium. However, in the presence of membranes containing acidic phospholipids, PKC bound at least eight calcium ions per protein. The presence of 1 microM phorbol dibutyrate (PDBu) in the dialysis buffer had little effect on these calcium-binding properties. Analysis of PKC-calcium binding by gel filtration under equilibrium conditions gave similar results; only membrane-associated PKC bound significant amounts of calcium. Consequently, PKC is a member of what may be a large group of proteins that bind calcium in a phospholipid-dependent manner. The calcium concentrations needed to induce PKC-membrane binding were similar to those needed for calcium binding (about 40 microM calcium at the midpoint). However, the calcium concentration required for PKC-membrane binding was strongly influenced by the phosphatidylserine composition of the membranes. Membranes with higher percentages of phosphatidylserine required lower concentrations of calcium. These properties suggested that the calcium sites may be generated at the interface between PKC and the membrane. Calcium may function as a bridge between PKC and phospholipids. These studies also suggested that calcium-dependent PKC-membrane binding and PKC function could be regulated by a number of factors in addition to calcium levels and diacylglycerol content of the membrane.

Effects of acclimation temperature on thermal tolerance and membrane phospholipid composition in the fruit fly Drosophila melanogaster

Czech Academy of Sciences Publication Activity Database

Overgaard, J.; Tomčala, Aleš; Sorensen, J. G.; Holmstrup, M.; Krogh, P. H.; Šimek, Petr; Košťál, Vladimír

2008-01-01

Roč. 54, č. 3 (2008), s. 619-629 ISSN 0022-1910 R&D Projects: GA ČR GA206/07/0269 Institutional research plan: CEZ:AV0Z50070508 Keywords : homeoviscous adaptation * membrane phospholipid * PE Subject RIV: ED - Physiology Impact factor: 2.155, year: 2008

Cholesterol autoxidation in phospholipid membrane bilayers

International Nuclear Information System (INIS)

Sevanian, A.; McLeod, L.L.

1987-01-01

Lipid peroxidation in unilamellar liposomes of known cholesterol-phospholipid composition was monitored under conditions of autoxidation or as induced by a superoxide radical generating system, gamma-irradiation or cumene hydroperoxide. Formation of cholesterol oxidation products was indexed to the level of lipid peroxidation. The major cholesterol oxidation products identified were 7-keto-cholesterol, isomeric cholesterol 5,6-epoxides, isomeric 7-hydroperoxides and isomeric 3,7-cholestane diols. Other commonly encountered products included 3,5-cholestadiene-7-one and cholestane-3 beta, 5 alpha, 6 beta-triol. Superoxide-dependent peroxidation required iron and produced a gradual increase in 7-keto-cholesterol and cholesterol epoxides. Cholesterol oxidation was greatest in liposomes containing high proportions of unsaturated phospholipid to cholesterol (4:1 molar ratio), intermediate with low phospholipid to cholesterol ratios (2:1) and least in liposomes prepared with dipalmitoylphosphatidylcholine and cholesterol. This relationship held regardless of the oxidizing conditions used. Cumene hydroperoxide-dependent lipid peroxidation and/or more prolonged oxidations with other oxidizing systems yielded a variety of products where cholesterol-5 beta,6 beta-epoxide, 7-ketocholesterol and the 7-hydroperoxides were most consistently elevated. Oxyradical initiation of lipid peroxidation produced a pattern of cholesterol oxidation products distinguishable from the pattern derived by cumene hydroperoxide-dependent peroxidation

Cortical actin networks induce spatio-temporal confinement of phospholipids in the plasma membrane--a minimally invasive investigation by STED-FCS.

Science.gov (United States)

Andrade, Débora M; Clausen, Mathias P; Keller, Jan; Mueller, Veronika; Wu, Congying; Bear, James E; Hell, Stefan W; Lagerholm, B Christoffer; Eggeling, Christian

2015-06-29

Important discoveries in the last decades have changed our view of the plasma membrane organisation. Specifically, the cortical cytoskeleton has emerged as a key modulator of the lateral diffusion of membrane proteins. Cytoskeleton-dependent compartmentalised lipid diffusion has been proposed, but this concept remains controversial because this phenomenon has thus far only been observed with artefact-prone probes in combination with a single technique: single particle tracking. In this paper, we report the first direct observation of compartmentalised phospholipid diffusion in the plasma membrane of living cells using a minimally invasive, fluorescent dye labelled lipid analogue. These observations were made using optical STED nanoscopy in combination with fluorescence correlation spectroscopy (STED-FCS), a technique which allows the study of membrane dynamics on a sub-millisecond time-scale and with a spatial resolution of down to 40 nm. Specifically, we find that compartmentalised phospholipid diffusion depends on the cortical actin cytoskeleton, and that this constrained diffusion is directly dependent on the F-actin branching nucleator Arp2/3. These findings provide solid evidence that the Arp2/3-dependent cortical actin cytoskeleton plays a pivotal role in the dynamic organisation of the plasma membrane, potentially regulating fundamental cellular processes.

Yeast PAH1-encoded phosphatidate phosphatase controls the expression of CHO1-encoded phosphatidylserine synthase for membrane phospholipid synthesis.

Science.gov (United States)

Han, Gil-Soo; Carman, George M

2017-08-11

The PAH1 -encoded phosphatidate phosphatase (PAP), which catalyzes the committed step for the synthesis of triacylglycerol in Saccharomyces cerevisiae , exerts a negative regulatory effect on the level of phosphatidate used for the de novo synthesis of membrane phospholipids. This raises the question whether PAP thereby affects the expression and activity of enzymes involved in phospholipid synthesis. Here, we examined the PAP-mediated regulation of CHO1 -encoded phosphatidylserine synthase (PSS), which catalyzes the committed step for the synthesis of major phospholipids via the CDP-diacylglycerol pathway. The lack of PAP in the pah1 Δ mutant highly elevated PSS activity, exhibiting a growth-dependent up-regulation from the exponential to the stationary phase of growth. Immunoblot analysis showed that the elevation of PSS activity results from an increase in the level of the enzyme encoded by CHO1 Truncation analysis and site-directed mutagenesis of the CHO1 promoter indicated that Cho1 expression in the pah1 Δ mutant is induced through the inositol-sensitive upstream activation sequence (UAS INO ), a cis -acting element for the phosphatidate-controlled Henry (Ino2-Ino4/Opi1) regulatory circuit. The abrogation of Cho1 induction and PSS activity by a CHO1 UAS INO mutation suppressed pah1 Δ effects on lipid synthesis, nuclear/endoplasmic reticulum membrane morphology, and lipid droplet formation, but not on growth at elevated temperature. Loss of the DGK1 -encoded diacylglycerol kinase, which converts diacylglycerol to phosphatidate, partially suppressed the pah1 Δ-mediated induction of Cho1 and PSS activity. Collectively, these data showed that PAP activity controls the expression of PSS for membrane phospholipid synthesis. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Efficient replacement of plasma membrane outer leaflet phospholipids and sphingolipids in cells with exogenous lipids.

Science.gov (United States)

Li, Guangtao; Kim, JiHyun; Huang, Zhen; St Clair, Johnna R; Brown, Deborah A; London, Erwin

2016-12-06

Our understanding of membranes and membrane lipid function has lagged far behind that of nucleic acids and proteins, largely because it is difficult to manipulate cellular membrane lipid composition. To help solve this problem, we show that methyl-α-cyclodextrin (MαCD)-catalyzed lipid exchange can be used to maximally replace the sphingolipids and phospholipids in the outer leaflet of the plasma membrane of living mammalian cells with exogenous lipids, including unnatural lipids. In addition, lipid exchange experiments revealed that 70-80% of cell sphingomyelin resided in the plasma membrane outer leaflet; the asymmetry of metabolically active cells was similar to that previously defined for erythrocytes, as judged by outer leaflet lipid composition; and plasma membrane outer leaflet phosphatidylcholine had a significantly lower level of unsaturation than phosphatidylcholine in the remainder of the cell. The data also provided a rough estimate for the total cellular lipids residing in the plasma membrane (about half). In addition to such lipidomics applications, the exchange method should have wide potential for investigations of lipid function and modification of cellular behavior by modification of lipids.

Mitochondrial cardiolipin/phospholipid trafficking: the role of membrane contact site complexes and lipid transfer proteins.

Science.gov (United States)

Schlattner, Uwe; Tokarska-Schlattner, Malgorzata; Rousseau, Denis; Boissan, Mathieu; Mannella, Carmen; Epand, Richard; Lacombe, Marie-Lise

2014-04-01

Historically, cellular trafficking of lipids has received much less attention than protein trafficking, mostly because its biological importance was underestimated, involved sorting and translocation mechanisms were not known, and analytical tools were limiting. This has changed during the last decade, and we discuss here some progress made in respect to mitochondria and the trafficking of phospholipids, in particular cardiolipin. Different membrane contact site or junction complexes and putative lipid transfer proteins for intra- and intermembrane lipid translocation have been described, involving mitochondrial inner and outer membrane, and the adjacent membranes of the endoplasmic reticulum. An image emerges how cardiolipin precursors, remodeling intermediates, mature cardiolipin and its oxidation products could migrate between membranes, and how this trafficking is involved in cardiolipin biosynthesis and cell signaling events. Particular emphasis in this review is given to mitochondrial nucleoside diphosphate kinase D and mitochondrial creatine kinases, which emerge to have roles in both, membrane junction formation and lipid transfer. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

Deformation of phospholipid vesicles in an optical stretcher

OpenAIRE

Delabre , Ulysse; Feld , Kasper; Crespo , Eleonore; Whyte , Graeme; Sykes , Cecile; Seifert , Udo; Guck , Jochen

2015-01-01

International audience; Phospholipid vesicles are common model systems for cell membranes. Important aspects of the membrane function relate to its mechanical properties. Here we have investigated the deformation behaviour of phospholipid vesicles in a dual-beam laser trap, also called an optical stretcher. This study explicitly makes use of the inherent heating present in such traps to investigate the dependence of vesicle deformation on temperature. By using lasers with different wavelength...

Evaluation of Ultrafiltration Performance for Phospholipid Separation

Science.gov (United States)

Aryanti, N.; Wardhani, D. H.; Maulana, Z. S.; Roberto, D.

2017-11-01

Ultrafiltration membrane for degumming of crude palm oil has been applied as an alternative method since the membrane process required less procedure than the conventional degumming. This research focused on the examination of ultrafiltration performance for phospholipid separation from model crude palm oil degumming. Specifically, profile flux and rejection, as well as blocking mechanism, were investigated. Feed consisting of Refined Crude Palm Oil - Isopropanol - Lecithin mixtures were represented as crude palm oil degumming. Lecithin was denoted a phospholipid component, and the concentrations of lecithin in feed were varied to 0.1%, 0.2%, and 0.3%. The concentration of phospholipid was determined as phosphor content. At the concentration of lecithin in feed representing phospholipid concentration of 8,45 mg/kg, 8,45 mg/kg, 24,87 mg/kg and 57,58 mg/kg, respectively. Flux profiles confirmed that there was a flux decline during filtration. In addition, the lecithin concentrations do not significantly effect on further flux decline. Rejection characteristic and phospholipid concentration in the permeate showed that the phospholipid rejections by ultrafiltration were in the range of 23-79,5% representing permeate’s phospholipid concentration of 1,73 - 44,25 mg/kg. Evaluation of fouling mechanism by Hermia’s blocking model confirmed that the standard blocking is the dominant mechanism in the ultrafiltration of lecithin mixture.

Annexin-Phospholipid Interactions. Functional Implications

Directory of Open Access Journals (Sweden)

Javier Turnay

2013-01-01

Full Text Available Annexins constitute an evolutionary conserved multigene protein superfamily characterized by their ability to interact with biological membranes in a calcium dependent manner. They are expressed by all living organisms with the exception of certain unicellular organisms. The vertebrate annexin core is composed of four (eight in annexin A6 homologous domains of around 70 amino acids, with the overall shape of a slightly bent ring surrounding a central hydrophilic pore. Calcium- and phospholipid-binding sites are located on the convex side while the N-terminus links domains I and IV on the concave side. The N-terminus region shows great variability in length and amino acid sequence and it greatly influences protein stability and specific functions of annexins. These proteins interact mainly with acidic phospholipids, such as phosphatidylserine, but differences are found regarding their affinity for lipids and calcium requirements for the interaction. Annexins are involved in a wide range of intra- and extracellular biological processes in vitro, most of them directly related with the conserved ability to bind to phospholipid bilayers: membrane trafficking, membrane-cytoskeleton anchorage, ion channel activity and regulation, as well as antiinflammatory and anticoagulant activities. However, the in vivo physiological functions of annexins are just beginning to be established.

Cell signalling and phospholipid metabolism

Energy Technology Data Exchange (ETDEWEB)

Boss, W.F.

1990-01-01

These studies explored whether phosphoinositide (PI) has a role in plants analogous to its role in animal cells. Although no parallel activity of PI in signal transduction was found in plant cells, activity of inositol phospholipid kinase was found to be modulated by light and by cell wall degrading enzymes. These studies indicate a major role for inositol phospholipids in plant growth and development as membrane effectors but not as a source of second messengers.

Co-existence of Gel and Fluid Lipid Domains in Single-component Phospholipid Membranes

Energy Technology Data Exchange (ETDEWEB)

Armstrong, Clare L [McMaster University; Barrett, M [McMaster University; Toppozini, L [McMaster University; Yamani, Zahra [Canadian Neutron Beam Centre, National Research Council, Chalk River Laboratorie; Kucerka, Norbert [Canadian Neutron Beam Centre and Comelius University (Slovakia); Katsaras, John [ORNL; Fragneto, Giovanna [Institut Laue-Langevin (ILL); Rheinstadter, Maikel C [McMaster University

2012-01-01

Lateral nanostructures in membranes, so-called rafts, are believed to strongly influence membrane properties and functions. The experimental observation of rafts has proven difficult as they are thought to be dynamic structures that likely fluctuate on nano- to microsecond time scales. Using neutron diffraction we present direct experimental evidence for the co-existence of gel and fluid lipid domains in a single-component phospholipid membrane made of DPPC as it undergoes its main phase transition. The coherence length of the neutron beam sets a lower limit for the size of structures that can be observed. Neutron coherence lengths between 30 and 242A used in this study were obtained by varying the incident neutron energy and the resolution of the neutron spectrometer. We observe Bragg peaks corresponding to co-existing nanometer sized structures, both in out-of-plane and in-plane scans, by tuning the neutron coherence length. During the main phase transition, instead of a continuous transition that shows a pseudo-critical behavior, we observe the co-existence of gel and fluid domains.

Eccentric contractions affect muscle membrane phospholipid fatty acid composition in rats

DEFF Research Database (Denmark)

Helge, Jørn Wulff; Therkildsen, K J; Jørgensen, T B

2001-01-01

This study investigated if prior eccentric contractions, and thus mechanical strain and muscle damage, exert an effect on the muscle membrane phospholipid fatty acid composition in rats, and whether a possible effect could be attenuated by dietary supplements. Twenty-three rats were randomised...... muscle, was excised from both legs. In the muscles stimulated to contract eccentrically, compared to the control muscles, the proportion of arachidonic acid, C20:4,n-6 (17.7 +/- 0.6; 16.4 +/- 0.4% of total fatty acids, respectively) and docosapentanoeic acid, C22:5,n-3 (2.9 +/- 0.1 and 2.7 +/- 0.......1% of total fatty acids, respectively) was uniformly higher across groups (P fatty acids) compared to the control leg (38.2 +/- 0...

Daptomycin resistance in enterococci is associated with distinct alterations of cell membrane phospholipid content.

Directory of Open Access Journals (Sweden)

Nagendra N Mishra

Full Text Available The lipopeptide antibiotic, daptomycin (DAP interacts with the bacterial cell membrane (CM. Development of DAP resistance during therapy in a clinical strain of Enterococcus faecalis was associated with mutations in genes encoding enzymes involved in cell envelope homeostasis and phospholipid metabolism. Here we characterized changes in CM phospholipid profiles associated with development of DAP resistance in clinical enterococcal strains.Using two clinical strain-pairs of DAP-susceptible and DAP-resistant E. faecalis (S613 vs. R712 and E. faecium (S447 vs. R446 recovered before and after DAP therapy, we compared four distinct CM profiles: phospholipid content, fatty acid composition, membrane fluidity and capacity to be permeabilized and/or depolarized by DAP. Additionally, we characterized the cell envelope of the E. faecium strain-pair by transmission electron microscopy and determined the relative cell surface charge of both strain-pairs.Both E. faecalis and E. faecium mainly contained four major CM PLs: phosphatidylglycerol (PG, cardiolipin, lysyl-phosphatidylglycerol (L-PG and glycerolphospho-diglycodiacylglycerol (GP-DGDAG. In addition, E. faecalis CMs (but not E. faecium also contained: i phosphatidic acid; and ii two other unknown species of amino-containing PLs. Development of DAP resistance in both enterococcal species was associated with a significant decrease in CM fluidity and PG content, with a concomitant increase in GP-DGDAG. The strain-pairs did not differ in their outer CM translocation (flipping of amino-containing PLs. Fatty acid content did not change in the E. faecalis strain-pair, whereas a significant decrease in unsaturated fatty acids was observed in the DAP-resistant E. faecium isolate R446 (vs S447. Resistance to DAP in E. faecium was associated with distinct structural alterations of the cell envelope and cell wall thickening, as well as a decreased ability of DAP to depolarize and permeabilize the CM

Characterization of Hydrophobic Interactions of Polymers with Water and Phospholipid Membranes Using Molecular Dynamics Simulations

Science.gov (United States)

Drenscko, Mihaela

small membranes using all atomistic and coarse-grained methods. The molecular interaction between common polymer chains used in biomedical applications and the cell membrane is unknown. This interaction may affect the biocompatibility of the polymer chains. Molecular dynamics simulations offer an emerging tool to characterize the interaction between common degradable polymer chains used in biomedical applications, such as polycaprolactone, and model cell membranes. We systematically characterize with long-time all-atomistic molecular dynamics simulations the interaction between single polycaprolactone chains of varying chain lengths with a model phospholipid membrane. We find that the length of polymer chain greatly affects the nature of interaction with the membrane, as well as the membrane properties. Furthermore, we next utilize advanced sampling techniques in molecular dynamics to characterize the two-dimensional free energy surface for the interaction of varying polymer chain lengths (short, intermediate, and long) with model cell membranes. We find that the free energy minimum shifts from the membrane-water interface to the hydrophobic core of the phospholipid membrane as a function of chain length. These results can be used to design polymer chain lengths and chemistries to optimize their interaction with cell membranes at the molecular level.

Covalent attachment of phospholipid analogous polymers to modify a polymeric membrane surface: a novel approach.

Science.gov (United States)

Xu, Zhi-Kang; Dai, Qing-Wen; Wu, Jian; Huang, Xiao-Jun; Yang, Qian

2004-02-17

A novel method for the surface modification of a microporous polypropylene membrane by tethering phospholipid analogous polymers (PAPs) is given, which includes the photoinduced graft polymerization of N,N-dimethylaminoethyl methacrylate (DMAEMA) and the ring-opening reaction of grafted poly-(DMAEMA) with 2-alkyloxy-2-oxo-1,3,2-dioxaphospholanes. Five 2-alkyloxy-2-oxo-1,3,2-dioxaphospholanes, containing octyloxy, dodecyloxy, tetradecyloxy, hexadecyloxy, and octadecyloxy groups in the molecular structure, were used to fabricate the PAP-modified polypropylene membranes. The attenuated total reflectance FT-IR spectra of the original, poly(DMAEMA)-grafted, and PAP-modified membranes confirmed the chemical changes on the membrane surface. Scanning electron microscope pictures showed that, compared with the original membrane, the surface porosities ofpoly(DMAEMA)-grafted and PAP-modified membranes were somewhat reduced. Water contact angles measured by the sessile drop method on PAP-modified membranes were slightly lower than that on the original polypropylene membrane, but higher than those on poly(DMAEMA)-grafted membranes with the exception of octyloxy-containing PAP-modified membranes. However, BSA adsorption experiments indicated that the five PAP-modified membranes had a much better protein-resistant property than the original polypropylene membrane and the poly(DMAEMA)-grafted membranes. For hexadecyloxy- and octadecyloxy-containing PAP-modified membranes, almost no protein adsorption was observed when the grafting degree was above 6 wt %. It was also found that the platelet adhesion was remarkably suppressed on the PAP-modified membranes. All these results demonstrate that the described approach is an effective way to improve the surface biocompatibility for polymeric membranes.

Plasma and erythrocyte membrane phospholipids and fatty acids in Italian general population and hemodialysis patients.

Science.gov (United States)

Dessì, Mariarita; Noce, Annalisa; Bertucci, Pierfrancesco; Noce, Gianluca; Rizza, Stefano; De Stefano, Alessandro; Manca di Villahermosa, Simone; Bernardini, Sergio; De Lorenzo, Antonino; Di Daniele, Nicola

2014-03-21

Dyslipidemia and abnormal phospholipid metabolism are frequent in uremic patients and increase their risk of cardiovascular disease (CVD): ω-3 polyunsaturated fatty acids (PUFAs) may reduce this risk in the general population. In this study we compared the plasma and erythrocyte cell membrane composition of PUFAs in a group of Caucasian hemodialysis (HD) patients and in a control group of healthy subjects and evaluated the erythrocyte/cell membrane fatty acid ratio as a marker of the dietary intake of phospholipids. The relationship between ω-3 and ω-6 fatty acids and the possible differences in PUFAs concentrations were also investigated. After obtaining a fully informed consent, a total of ninety-nine HD patients and 160 non uremic control subjects from "Tor Vergata" University Hospital were enrolled into the study. None of them took antioxidant drugs or dietary supplements for at least 90 days prior to the observation. Blood samples were analysed by gas-chromatographic coupled to a mass spectrometric detector.The daily intake of total calories, proteins, lipids and carbohydrates is significantly lower in HD patients than in controls (p HD patients (p HD patients, due to the removal of nutrients during the dialysis and to persistent malnutrition. A dietary treatment addressed to increase plasma ω-3 PUFAs and to optimize ω-6/ω-3 ratio may exert a protective action and reduce the risk of CVD in HD patient.

Phospholipase A2 activity-dependent and -independent fusogenic activity of Naja nigricollis CMS-9 on zwitterionic and anionic phospholipid vesicles.

Science.gov (United States)

Chiou, Yi-Ling; Chen, Ying-Jung; Lin, Shinne-Ren; Chang, Long-Sen

2011-11-01

CMS-9, a phospholipase A(2) (PLA(2)) from Naja nigricollis venom, induced the death of human breast cancer MCF-7 cells accompanied with the formation of cell clumps without clear boundaries between cells. Annexin V-FITC staining indicated that abundant phosphatidylserine appeared on the outer membrane of MCF-7 cell clumps, implying the possibility that CMS-9 may promote membrane fusion via anionic phospholipids. To validate this proposition, fusogenic activity of CMS-9 on vesicles composed of zwitterionic phospholipid alone or a combination of zwitterionic and anionic phospholipids was examined. Although CMS-9-induced fusion of zwitterionic phospholipid vesicles depended on PLA(2) activity, CMS-9-induced fusion of vesicles containing anionic phospholipids could occur without the involvement of PLA(2) activity. Membrane-damaging activity of CMS-9 was associated with its fusogenicity. Moreover, CMS-9 induced differently membrane leakage and membrane fusion of vesicles with different compositions. Membrane fluidity and binding capability with phospholipid vesicles were not related to the fusogenicity of CMS-9. However, membrane-bound conformation and mode of CMS-9 depended on phospholipid compositions. Collectively, our data suggest that PLA(2) activity-dependent and -independent fusogenicity of CMS-9 are closely related to its membrane-bound modes and targeted membrane compositions. Copyright © 2011 Elsevier Ltd. All rights reserved.

Promiscuous and specific phospholipid binding by domains in ZAC, a membrane-associated Arabidopsis protein with an ARF GAP zinc finger and a C2 domain

DEFF Research Database (Denmark)

Jensen, R B; Lykke-Andersen, K; Frandsen, G I

2000-01-01

domain are separated by a region without homology to other known proteins. Zac promoter/beta-glucuronidase reporter assays revealed highest expression levels in flowering tissue, rosettes and roots. ZAC protein was immuno-detected mainly in association with membranes and fractionated with Golgi...... and plasma membrane marker proteins. ZAC membrane association was confirmed in assays by a fusion between ZAC and the green fluorescence protein and prompted an analysis of the in vitro phospholipid-binding ability of ZAC. Phospholipid dot-blot and liposome-binding assays indicated that fusion proteins...... zinc finger motif, but proteins containing only the zinc finger domain (residues 1-105) did not bind PI-3-P. Recombinant ZAC possessed GTPase-activating activity on Arabidopsis ARF proteins. These data identify a novel PI-3-P-binding protein region and thereby provide evidence...

A conserved endoplasmic reticulum membrane protein complex (EMC facilitates phospholipid transfer from the ER to mitochondria.

Directory of Open Access Journals (Sweden)

Sujoy Lahiri

2014-10-01

Full Text Available Mitochondrial membrane biogenesis and lipid metabolism require phospholipid transfer from the endoplasmic reticulum (ER to mitochondria. Transfer is thought to occur at regions of close contact of these organelles and to be nonvesicular, but the mechanism is not known. Here we used a novel genetic screen in S. cerevisiae to identify mutants with defects in lipid exchange between the ER and mitochondria. We show that a strain missing multiple components of the conserved ER membrane protein complex (EMC has decreased phosphatidylserine (PS transfer from the ER to mitochondria. Mitochondria from this strain have significantly reduced levels of PS and its derivative phosphatidylethanolamine (PE. Cells lacking EMC proteins and the ER-mitochondria tethering complex called ERMES (the ER-mitochondria encounter structure are inviable, suggesting that the EMC also functions as a tether. These defects are corrected by expression of an engineered ER-mitochondrial tethering protein that artificially tethers the ER to mitochondria. EMC mutants have a significant reduction in the amount of ER tethered to mitochondria even though ERMES remained intact in these mutants, suggesting that the EMC performs an additional tethering function to ERMES. We find that all Emc proteins interact with the mitochondrial translocase of the outer membrane (TOM complex protein Tom5 and this interaction is important for PS transfer and cell growth, suggesting that the EMC forms a tether by associating with the TOM complex. Together, our findings support that the EMC tethers ER to mitochondria, which is required for phospholipid synthesis and cell growth.

Harvesting Water from Air: Using Anhydrous Salt with Sunlight

KAUST Repository

Li, Renyuan

2018-04-02

Atmospheric water is abundant alternative water resource, equivalent to 6 times of water in all rivers on Earth. This work screens 14 common anhydrous and hydrated salt couples in terms of their physical and chemical stability, water vapor harvesting and release capacity under relevant application scenarios. Among the salts screened, copper chloride (CuCl2), copper sulfate (CuSO4) and magnesium sulfate (MgSO4) distinguish themselves and are further made into bi-layer water collection devices, with the top layer being photothermal layer while the bottom layer being salt-loaded fibrous membrane. The water collection devices are capable of capturing water vapor out of the air with low relative humidity (down to 15 %) and releasing water under regular and even weakened sunlight (i.e. 0.7 kW/m2). The work shines light on the potential use of anhydrous salt towards producing drinking water in water scarce regions.

Autistic disorder and phospholipids: A review.

Science.gov (United States)

Brown, Christine M; Austin, David W

2011-01-01

Dysregulated phospholipid metabolism has been proposed as an underlying biological component of neurodevelopmental disorders such as autistic disorder (AD) and attention-deficit/hyperactivity disorder (ADHD). This review provides an overview of fatty acid and phospholipid metabolism and evidence for phospholipid dysregulation with reference to the membrane hypothesis of schizophrenia. While there is evidence that phospholipid metabolism is at least impaired in individuals with AD, it has not been established whether phospholipid metabolism is implicated in causal, mechanistic or epiphenomenological models. More research is needed to ascertain whether breastfeeding, and specifically, the administration of colostrum or an adequate substitute can play a preventative role by supplying the neonate with essential fatty acids (EFAs) at a critical juncture in their development. Regarding treatment, further clinical trials of EFA supplementation are essential to determine the efficacy of EFAs in reducing AD symptomatology and whether supplementation can serve as a cost-effective and readily available intervention. Crown Copyright © 2010. Published by Elsevier Ltd. All rights reserved.

Structural Interpretation of the Large Slowdown of Water Dynamics at Stacked Phospholipid Membranes for Decreasing Hydration Level: All-Atom Molecular Dynamics

Directory of Open Access Journals (Sweden)

Carles Calero

2016-04-01

Full Text Available Hydration water determines the stability and function of phospholipid membranes as well as the interaction of membranes with other molecules. Experiments and simulations have shown that water dynamics slows down dramatically as the hydration decreases, suggesting that the interfacial water that dominates the average dynamics at low hydration is slower than water away from the membrane. Here, based on all-atom molecular dynamics simulations, we provide an interpretation of the slowdown of interfacial water in terms of the structure and dynamics of water–water and water–lipid hydrogen bonds (HBs. We calculate the rotational and translational slowdown of the dynamics of water confined in stacked phospholipid membranes at different levels of hydration, from completely hydrated to poorly hydrated membranes. For all hydrations, we analyze the distribution of HBs and find that water–lipids HBs last longer than water–water HBs and that at low hydration most of the water is in the interior of the membrane. We also show that water–water HBs become more persistent as the hydration is lowered. We attribute this effect (i to HBs between water molecules that form, in turn, persistent HBs with lipids; (ii to the hindering of the H-bonding switching between water molecules due to the lower water density at the interface; and (iii to the higher probability of water–lipid HBs as the hydration decreases. Our interpretation of the large dynamic slowdown in water under dehydration is potentially relevant in understanding membrane biophysics at different hydration levels.

21 CFR 168.110 - Dextrose anhydrous.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Dextrose anhydrous. 168.110 Section 168.110 Food... Table Sirups § 168.110 Dextrose anhydrous. (a) Dextrose anhydrous is purified and crystallized D-glucose... solids content is not less than 98.0 percent m/m. (b) The name of the food is “Dextrose anhydrous” or...

Composite polymer electrolyte containing ionic liquid and functionalized polyhedral oligomeric silsesquioxanes for anhydrous PEM applications.

Science.gov (United States)

Subianto, Surya; Mistry, Mayur K; Choudhury, Namita Roy; Dutta, Naba K; Knott, Robert

2009-06-01

A new type of supported liquid membrane was made by combining an ionic liquid (IL) with a Nafion membrane reinforced with multifunctional polyhedral oligomeric silsesquioxanes (POSSs) using a layer-by-layer strategy for anhydrous proton-exchange membrane (PEM) application. The POSS was functionalized by direct sulfonation, and the sulfonated POSS (S-POSS) was incorporated into Nafion 117 membranes by the infiltration method. The resultant hybrid membrane shows strong ionic interaction between the Nafion matrix and the multifunctional POSS, resulting in increased glass transition temperature and thermal stability at very low loadings of S-POSS (1%). The presence of S-POSS has also improved the proton conductivity especially at low humidities, where it shows a marked increase due to its confinement in the ionic domains and promotes water uptake by capillary condensation. In order to achieve anhydrous conductivity, the IL 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide (BMI-BTSI) was incorporated into these membranes to provide proton conduction in the absence of water. Although the incorporation of an IL shows a plasticizing effect on the Nafion membrane, the S-POSS composite membrane with an IL shows a higher modulus at high temperatures compared to Nafion 117 and a Nafion-IL membrane, with significantly higher proton conductivity (5 mS/cm at 150 degrees C with 20% IL). This shows the ability of the multifunctional POSS and IL to work symbiotically to achieve the desirable proton conductivity and mechanical properties of such membranes by enhancing the ionic interaction within the material.

Supercritical CO2 induces marked changes in membrane phospholipids composition in Escherichia coli K12.

Science.gov (United States)

Tamburini, Sabrina; Anesi, Andrea; Ferrentino, Giovanna; Spilimbergo, Sara; Guella, Graziano; Jousson, Olivier

2014-06-01

Supercritical carbon dioxide (SC-CO2) treatment is one of the most promising alternative techniques for pasteurization of both liquid and solid food products. The inhibitory effect of SC-CO2 on bacterial growth has been investigated in different species, but the precise mechanism of action remains unknown. Membrane permeabilization has been proposed to be the first event in SC-CO2-mediated inactivation. Flow cytometry, high performance liquid chromatography–electrospray ionization–mass spectrometry and NMR analyses were performed to investigate the effect of SC-CO2 treatment on membrane lipid profile and membrane permeability in Escherichia coli K12. After 15 min of SC-CO2 treatment at 120 bar and 35 °C, the majority of bacterial cells dissipated their membrane potential (95 %) and lost membrane integrity, as 81 % become partially permeabilized and 18 % fully permeabilized. Membrane permeabilization was associated with a 20 % decrease in bacterial biovolume and to a strong (>50 %) reduction in phosphatidylglycerol (PG) membrane lipids, without altering the fatty acid composition and the degree of unsaturation of acyl chains. PGs are thought to play an important role in membrane stability, by reducing motion of phosphatidylethanolamine (PE) along the membrane bilayer, therefore promoting the formation of inter-lipid hydrogen bonds. In addition, the decrease in intracellular pH induced by SC-CO2 likely alters the chemical properties of phospholipids and the PE/PG ratio. Biophysical effects of SC-CO2 thus cause a strong perturbation of membrane architecture in E. coli, and such alterations are likely associated with its strong inactivation effect.

Characterization of Phospholipids in Insulin Secretory Granules and Mitochondria in Pancreatic Beta Cells and Their Changes with Glucose Stimulation*

Science.gov (United States)

MacDonald, Michael J.; Ade, Lacmbouh; Ntambi, James M.; Ansari, Israr-Ul H.; Stoker, Scott W.

2015-01-01

The lipid composition of insulin secretory granules (ISG) has never previously been thoroughly characterized. We characterized the phospholipid composition of ISG and mitochondria in pancreatic beta cells without and with glucose stimulation. The phospholipid/protein ratios of most phospholipids containing unsaturated fatty acids were higher in ISG than in whole cells and in mitochondria. The concentrations of negatively charged phospholipids, phosphatidylserine, and phosphatidylinositol in ISG were 5-fold higher than in the whole cell. In ISG phosphatidylserine, phosphatidylinositol, phosphatidylethanolamine, and sphingomyelin, fatty acids 12:0 and 14:0 were high, as were phosphatidylserine and phosphatidylinositol containing 18-carbon unsaturated FA. With glucose stimulation, the concentration of many ISG phosphatidylserines and phosphatidylinositols increased; unsaturated fatty acids in phosphatidylserine increased; and most phosphatidylethanolamines, phosphatidylcholines, sphingomyelins, and lysophosphatidylcholines were unchanged. Unsaturation and shorter fatty acid length in phospholipids facilitate curvature and fluidity of membranes, which favors fusion of membranes. Recent evidence suggests that negatively charged phospholipids, such as phosphatidylserine, act as coupling factors enhancing the interaction of positively charged regions in SNARE proteins in synaptic or secretory vesicle membrane lipid bilayers with positively charged regions in SNARE proteins in the plasma membrane lipid bilayer to facilitate docking of vesicles to the plasma membrane during exocytosis. The results indicate that ISG phospholipids are in a dynamic state and are consistent with the idea that changes in ISG phospholipids facilitate fusion of ISG with the plasma membrane-enhancing glucose-stimulated insulin exocytosis. PMID:25762724

Formation and Regulation of Mitochondrial Membranes

Directory of Open Access Journals (Sweden)

Laila Cigana Schenkel

2014-01-01

Full Text Available Mitochondrial membrane phospholipids are essential for the mitochondrial architecture, the activity of respiratory proteins, and the transport of proteins into the mitochondria. The accumulation of phospholipids within mitochondria depends on a coordinate synthesis, degradation, and trafficking of phospholipids between the endoplasmic reticulum (ER and mitochondria as well as intramitochondrial lipid trafficking. Several studies highlight the contribution of dietary fatty acids to the remodeling of phospholipids and mitochondrial membrane homeostasis. Understanding the role of phospholipids in the mitochondrial membrane and their metabolism will shed light on the molecular mechanisms involved in the regulation of mitochondrial function and in the mitochondrial-related diseases.

Cell signalling and phospholipid metabolism. Final report

Energy Technology Data Exchange (ETDEWEB)

Boss, W.F.

1990-12-31

These studies explored whether phosphoinositide (PI) has a role in plants analogous to its role in animal cells. Although no parallel activity of PI in signal transduction was found in plant cells, activity of inositol phospholipid kinase was found to be modulated by light and by cell wall degrading enzymes. These studies indicate a major role for inositol phospholipids in plant growth and development as membrane effectors but not as a source of second messengers.

Anhydrous proton conductor based on composites of PEO and ATMP

International Nuclear Information System (INIS)

Sun Baoying; Qiu Xinping; Zhu Wentao

2011-01-01

A new type anhydrous PEM material based on Poly (ethylene oxide) (PEO)/Amino Trimethylene Phosphonic Acid (ATMP) composite was prepared. In this study, PEO assumed to 'grab' protons via hydrogen bond between PEO and ATMP. Based on this point, the PEO/ATMP composites were prepared firstly as the preliminary study to verify this proton conducting system. Then, PVDF was added to enhance the membrane's stability. The PVDF/PEO/ATMP composite membranes were thermally stable up to 200 o C in the studied composition ranges. The membrane had relatively compact structure by SEM images. Proton conductivity of 59% PVDF/29% PEO/12% ATMP was up to 6.71 x 10 -3 S cm -1 at 86 o C after doping with 7.9 wt% phosphoric acid without extra humidification.

Membrane-surfactant interactions. The role of surfactant in mitochondrial complex III-phospholipid-Triton X-100 mixed micelles

International Nuclear Information System (INIS)

Valpuesta, J.M.; Arrondo, J.L.; Barbero, M.C.; Pons, M.; Goni, F.M.

1986-01-01

Complex III (ubiquinol-cytochrome c reductase) was purified from beef heart mitochondria in the form of protein-phospholipid-Triton X-100 mixed micelles (about 1:80:100 molar ratio). Detergent may be totally removed by sucrose density gradient centrifugation, and the resulting lipoprotein complexes retain full enzyme activity. In order to understand the role of surfactant in the mixed micelles, and the interaction of Triton X-100 with integral membrane proteins and phospholipid bilayers, both the protein-lipid-surfactant mixed micelles and the detergent-free lipoprotein system were examined from the point of view of particle size and ultrastructure, enzyme activity, tryptophan fluorescence quenching, 31P NMR, and Fourier transform infrared spectroscopy. The NMR and IR spectroscopic studies show that surfactant withdrawal induces a profound change in phospholipid architecture, from a micellar to a lamellar-like phase. However, electron microscopic observations fail to reveal the existence of lipid bilayers in the absence of detergent. We suggest that, under these conditions, the lipid:protein molar ratio (80:1) is too low to permit the formation of lipid bilayer planes, but the relative orientation and mobility of phospholipids with respect to proteins is similar to that of the lamellar phase. Protein conformational changes are also detected as a consequence of surfactant removal. Fourier transform infrared spectroscopy indicates an increase of peptide beta-structure in the absence of Triton X-100; changes in the amide II/amide I intensity ratio are also detected, although the precise meaning of these observations is unclear

13C- and 15N-Labeling Strategies Combined with Mass Spectrometry Comprehensively Quantify Phospholipid Dynamics in C. elegans.

Directory of Open Access Journals (Sweden)

Blair C R Dancy

Full Text Available Membranes define cellular and organelle boundaries, a function that is critical to all living systems. Like other biomolecules, membrane lipids are dynamically maintained, but current methods are extremely limited for monitoring lipid dynamics in living animals. We developed novel strategies in C. elegans combining 13C and 15N stable isotopes with mass spectrometry to directly quantify the replenishment rates of the individual fatty acids and intact phospholipids of the membrane. Using multiple measurements of phospholipid dynamics, we found that the phospholipid pools are replaced rapidly and at rates nearly double the turnover measured for neutral lipid populations. In fact, our analysis shows that the majority of membrane lipids are replaced each day. Furthermore, we found that stearoyl-CoA desaturases (SCDs, critical enzymes in polyunsaturated fatty acid production, play an unexpected role in influencing the overall rates of membrane maintenance as SCD depletion affected the turnover of nearly all membrane lipids. Additionally, the compromised membrane maintenance as defined by LC-MS/MS with SCD RNAi resulted in active phospholipid remodeling that we predict is critical to alleviate the impact of reduced membrane maintenance in these animals. Not only have these combined methodologies identified new facets of the impact of SCDs on the membrane, but they also have great potential to reveal many undiscovered regulators of phospholipid metabolism.

Characterization of phospholipids in insulin secretory granules and mitochondria in pancreatic beta cells and their changes with glucose stimulation.

Science.gov (United States)

MacDonald, Michael J; Ade, Lacmbouh; Ntambi, James M; Ansari, Israr-Ul H; Stoker, Scott W

2015-04-24

The lipid composition of insulin secretory granules (ISG) has never previously been thoroughly characterized. We characterized the phospholipid composition of ISG and mitochondria in pancreatic beta cells without and with glucose stimulation. The phospholipid/protein ratios of most phospholipids containing unsaturated fatty acids were higher in ISG than in whole cells and in mitochondria. The concentrations of negatively charged phospholipids, phosphatidylserine, and phosphatidylinositol in ISG were 5-fold higher than in the whole cell. In ISG phosphatidylserine, phosphatidylinositol, phosphatidylethanolamine, and sphingomyelin, fatty acids 12:0 and 14:0 were high, as were phosphatidylserine and phosphatidylinositol containing 18-carbon unsaturated FA. With glucose stimulation, the concentration of many ISG phosphatidylserines and phosphatidylinositols increased; unsaturated fatty acids in phosphatidylserine increased; and most phosphatidylethanolamines, phosphatidylcholines, sphingomyelins, and lysophosphatidylcholines were unchanged. Unsaturation and shorter fatty acid length in phospholipids facilitate curvature and fluidity of membranes, which favors fusion of membranes. Recent evidence suggests that negatively charged phospholipids, such as phosphatidylserine, act as coupling factors enhancing the interaction of positively charged regions in SNARE proteins in synaptic or secretory vesicle membrane lipid bilayers with positively charged regions in SNARE proteins in the plasma membrane lipid bilayer to facilitate docking of vesicles to the plasma membrane during exocytosis. The results indicate that ISG phospholipids are in a dynamic state and are consistent with the idea that changes in ISG phospholipids facilitate fusion of ISG with the plasma membrane-enhancing glucose-stimulated insulin exocytosis. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Novicidin interactions with phospholipid membranes

DEFF Research Database (Denmark)

Balakrishnan, Vijay Shankar

Antimicrobial peptides target bacterial cell membranes and are considered as potential antibiotics. Their interactions with cell membranes are studied using different approaches. This thesis comprises of the biophysical investigations on the antimicrobial peptide Novicidin, interacting with lipos......Antimicrobial peptides target bacterial cell membranes and are considered as potential antibiotics. Their interactions with cell membranes are studied using different approaches. This thesis comprises of the biophysical investigations on the antimicrobial peptide Novicidin, interacting...... with liposomes. The lipid-induced changes in the peptide due to membrane binding, and the peptide-induced changes in the membrane properties were investigated using various spectroscopic and calorimetric methods, and the structural and thermodynamic aspects of peptide-lipid interactions are discussed. This helps...

Neutron diffraction studies of amphipathic helices in phospholipid bilayers

Energy Technology Data Exchange (ETDEWEB)

Bradshaw, J.P.; Gilchrist, P.J. [Univ. of Edinburgh (United Kingdom); Duff, K.C. [Univ. of Edinburgh Medical School (United Kingdom); Saxena, A.M. [Brookhaven National Laboratory, Upton, NY (United States)

1994-12-31

The structural feature which is thought to facilitate the interaction of many peptides with phospholipid bilayers is the ability to fold into an amphipathic helix. In most cases the exact location and orientation of this helix with respect to the membrane is not known, and may vary with factors such as pH and phospholipid content of the bilayer. The growing interest in this area is stimulated by indications that similar interactions can contribute to the binding of certain hormones to their cell-surface receptors. We have been using the techniques of neutron diffraction from stacked phospholipid bilayers in an attempt to investigate this phenomenon with a number of membrane-active peptides. Here we report some of our findings with three of these: the bee venom melittin; the hormone calcitonin; and a synthetic peptide representing the ion channel fragment of influenza A M2 protein.

Neutron diffraction studies of amphipathic helices in phospholipid bilayers

International Nuclear Information System (INIS)

Bradshaw, J.P.; Gilchrist, P.J.; Duff, K.C.; Saxena, A.M.

1994-01-01

The structural feature which is thought to facilitate the interaction of many peptides with phospholipid bilayers is the ability to fold into an amphipathic helix. In most cases the exact location and orientation of this helix with respect to the membrane is not known, and may vary with factors such as pH and phospholipid content of the bilayer. The growing interest in this area is stimulated by indications that similar interactions can contribute to the binding of certain hormones to their cell-surface receptors. We have been using the techniques of neutron diffraction from stacked phospholipid bilayers in an attempt to investigate this phenomenon with a number of membrane-active peptides. Here we report some of our findings with three of these: the bee venom melittin; the hormone calcitonin; and a synthetic peptide representing the ion channel fragment of influenza A M2 protein

Anhydrous proton conductor based on composites of PEO and ATMP

Energy Technology Data Exchange (ETDEWEB)

Sun Baoying [Key Lab of Organic Optoelectronics and Molecular Engineering, Department of Chemistry, Tsinghua University, Beijing 100084 (China); Qiu Xinping, E-mail: qiuxp@tsinghua.edu.c [Key Lab of Organic Optoelectronics and Molecular Engineering, Department of Chemistry, Tsinghua University, Beijing 100084 (China); Zhu Wentao [Key Lab of Organic Optoelectronics and Molecular Engineering, Department of Chemistry, Tsinghua University, Beijing 100084 (China)

2011-04-15

A new type anhydrous PEM material based on Poly (ethylene oxide) (PEO)/Amino Trimethylene Phosphonic Acid (ATMP) composite was prepared. In this study, PEO assumed to 'grab' protons via hydrogen bond between PEO and ATMP. Based on this point, the PEO/ATMP composites were prepared firstly as the preliminary study to verify this proton conducting system. Then, PVDF was added to enhance the membrane's stability. The PVDF/PEO/ATMP composite membranes were thermally stable up to 200 {sup o}C in the studied composition ranges. The membrane had relatively compact structure by SEM images. Proton conductivity of 59% PVDF/29% PEO/12% ATMP was up to 6.71 x 10 {sup -3} S cm{sup -1} at 86 {sup o}C after doping with 7.9 wt% phosphoric acid without extra humidification.

Structure and mechanism of ATP-dependent phospholipid transporters

DEFF Research Database (Denmark)

Lopez Marques, Rosa Laura; Poulsen, Lisbeth Rosager; Bailly, Aurélien

2015-01-01

Background ATP-binding cassette (ABC) transporters and P4-ATPases are two large and seemingly unrelated families of primary active pumps involved in moving phospholipids from one leaflet of a biological membrane to the other. Scope of review This review aims to identify common mechanistic features...... in the way phospholipid flipping is carried out by two evolutionarily unrelated families of transporters. Major conclusions Both protein families hydrolyze ATP, although they employ different mechanisms to use it, and have a comparable size with twelve transmembrane segments in the functional unit. Further......, despite differences in overall architecture, both appear to operate by an alternating access mechanism and during transport they might allow access of phospholipids to the internal part of the transmembrane domain. The latter feature is obvious for ABC transporters, but phospholipids and other hydrophobic...

Binding of the radioprotective agent cysteamine with the phospholipidic membrane headgroup-interface region

Energy Technology Data Exchange (ETDEWEB)

Berleur, F; Roman, V; Jaskierowicz, D; Fatome, M; Leterrier, F; Ter-Minassian-Saraga, L; Madelmont, G

1985-09-01

The interaction of the aminothiol radioprotector cysteamine (..beta..-mercaptoethylamine)(CYST) with dipalmitoylphosphatidylcholine (DPPC) artificial membranes has been studied by differential scanning calorimetry (DSC), turbidimetry and spin labeling. This hydrophilic molecule displays a biphasic, concentration-dependent binding to the phospholipidic head groups at neutral pH. In the CYST/DPPC molar ratio 1:160-1:2 (mole/mole) an increasing ordering effect is observed. At high concentrations (over 3:1 ratio), this ordering effect decreases. With the symmetric disulfide dimer cystamine, the biphasic effect is not shown and the membrane rigidity decrease is obtained only at concentration ratio higher than 1:1. The charge repartition of the cysteamine molecule has been shown to be disymmetric, +0.52 e on the NH/sub 3/ group and +0.19 e on the SH extremity, whereas the cystamine molecule is electrostatically symmetrical. These properties could be related to their membrane effects. With cysteamine, at a low concentration, an electrostatic bridging between the negatively charged phosphate groups of the polar heads induces the increase in membrane stability: the molecules behave like a divalent cation. At high concentration a displacement of the slightly charged SH extremity by the amine disrupts the bridges and induces the decrease in rigidity: the drug behaves like a monovalent cation. Due to its symmetric charge and its double length, such an effect is not observed with cystamine. This study could bring further information about the interactions between cysteamine and polyelectrolytic structures (ADN for example) and about the radioprotective properties of this drug.

Morphological and Physical Analysis of Natural Phospholipids-Based Biomembranes

OpenAIRE

Jacquot, Adrien; Francius, Grégory; Razafitianamaharavo, Angelina; Dehghani, Fariba; Tamayol, Ali; Linder, Michel; Arab-Tehrany, Elmira

2014-01-01

International audience; Background: Liposomes are currently an important part of biological, pharmaceutical, medical and nutritional research, as they are considered to be among the most effective carriers for the introduction of various types of bioactive agents into target cells.Scope of Review: In this work, we study the lipid organization and mechanical properties of biomembranes made of marine and plant phospholipids. Membranes based on phospholipids extracted from rapeseed and salmon ar...

Optimization of transversal relaxation of nitroxides for pulsed electron-electron double resonance spectroscopy in phospholipid membranes.

Science.gov (United States)

Dastvan, Reza; Bode, Bela E; Karuppiah, Muruga Poopathi Raja; Marko, Andriy; Lyubenova, Sevdalina; Schwalbe, Harald; Prisner, Thomas F

2010-10-28

Pulsed electron-electron double resonance (PELDOR) spectroscopy is increasingly applied to spin-labeled membrane proteins. However, after reconstitution into liposomes, spin labels often exhibit a much faster transversal relaxation (T(m)) than in detergent micelles, thus limiting application of the method in lipid bilayers. In this study, the main reasons for enhanced transversal relaxation in phospholipid membranes were investigated systematically by use of spin-labeled derivatives of stearic acid and phosphatidylcholine as well as spin-labeled derivatives of the channel-forming peptide gramicidin A under the conditions typically employed for PELDOR distance measurements. Our results clearly show that dephasing due to instantaneous diffusion that depends on dipolar interaction among electron spins is an important contributor to the fast echo decay in cases of high local concentrations of spin labels in membranes. The main difference between spin labels in detergent micelles and membranes is their local concentration. Consequently, avoiding spin clustering and suppressing instantaneous diffusion is the key step for maximizing PELDOR sensitivity in lipid membranes. Even though proton spin diffusion is an important relaxation mechanism, only in samples of low local concentrations does deuteration of acyl chains and buffer significantly prolong T(m). In these cases, values of up to 7 μs have been achieved. Furthermore, our study revealed that membrane composition and labeling position in the membrane can also affect T(m), either by promoting the segregation of spin-labeled species or by altering their exposure to matrix protons. Effects of other experimental parameters including temperature (<50 K), presence of oxygen, and cryoprotectant type are negligible under our experimental conditions.

46 CFR 151.50-32 - Ammonia, anhydrous.

Science.gov (United States)

2010-10-01

... enclosed anhydrous ammonia tanks complies with the following chemical and physical properties: (1) Boiling... requirements of § 151.50-30 for compressed gases are also applicable to the shipment of anhydrous ammonia...

Aqueous magnesium as an environmental selection pressure in the evolution of phospholipid membranes on early earth

Science.gov (United States)

Dalai, Punam; Ustriyana, Putu; Sahai, Nita

2018-02-01

Early compartmentalization of simple biomolecules by membrane bilayers was, presumably, a critical step in the emergence of the first cell-like entities, protocells. Their membranes were likely composed of single chain amphiphiles (SCAs), but pure SCA membranes especially those with short-chains are highly unstable towards divalent cations, which are ubiquitous in aqueous environments. The prebiotic synthesis of phospholipids (PLs), even in only trace amounts, may also have been possible. PL membranes are much more stable towards divalent cations. Here, we show the transition of fatty acid membranes to mixed fatty acid-PL and, finally, to PL membranes in the presence of Mg2+, which acts as an environmental selection pressure, and we propose different mechanisms for the observed increased Mg2+-immunity. The "fatal" concentration ([Mg2+]fatal) at which vesicles are disrupted increased dramatically by an order of magnitude from OA to mixed to POPC vesicles. Two mechanisms for the increasing immunity were determined. The negative charge density of the vesicles decreased with increasing POPC content, so more Mg2+ was required for disruption. More interestingly, Mg2+ preferentially bound to and abstracted OA from mixed lipid membranes, resulting in relatively POPC-enriched vesicles compared to the initial ratio. The effect was the most dramatic for the largest initial OA-POPC ratio representing the most primitive protocells. Thus, Mg2+ acted to evolve the mixed membrane composition towards PL enrichment. To the best of our knowledge, this is the first report of selective lipid abstraction from mixed SCA-PL vesicles. These results may hold implications for accommodating prebiotic Mg2+-promoted processes such as non-enzymatic RNA polymerization on early Earth.

Biosynthesis of ether-phospholipids including plasmalogens, peroxisomes and human disease: new insights into an old problem

NARCIS (Netherlands)

Wanders, Ronald J. A.; Brites, Pedro

2010-01-01

Ether-phospholipids represent an important subclass of phospholipids in animal cell membranes characterized by the presence of an ether bond at the sn-I position and the enrichment of PUFAs at the sn-2 position. Of the different ether-phospholipids, plasmalogens are the most abundant form and their

46 CFR 98.25-5 - How anhydrous ammonia may be carried.

Science.gov (United States)

2010-10-01

... 46 Shipping 4 2010-10-01 2010-10-01 false How anhydrous ammonia may be carried. 98.25-5 Section 98... Anhydrous Ammonia in Bulk § 98.25-5 How anhydrous ammonia may be carried. (a) Anhydrous ammonia shall be..., except as otherwise provided in paragraph (b) of this section. (b) When anhydrous ammonia is to be...

Enhanced root growth in phosphate-starved Arabidopsis by stimulating de novo phospholipid biosynthesis through the overexpression of LYSOPHOSPHATIDIC ACID ACYLTRANSFERASE 2 (LPAT2).

Science.gov (United States)

Angkawijaya, Artik Elisa; Nguyen, Van Cam; Nakamura, Yuki

2017-09-01

Upon phosphate starvation, plants retard shoot growth but promote root development presumably to enhance phosphate assimilation from the ground. Membrane lipid remodelling is a metabolic adaptation that replaces membrane phospholipids by non-phosphorous galactolipids, thereby allowing plants to obtain scarce phosphate yet maintain the membrane structure. However, stoichiometry of this phospholipid-to-galactolipid conversion may not account for the massive demand of membrane lipids that enables active growth of roots under phosphate starvation, thereby suggesting the involvement of de novo phospholipid biosynthesis, which is not represented in the current model. We overexpressed an endoplasmic reticulum-localized lysophosphatidic acid acyltransferase, LPAT2, a key enzyme that catalyses the last step of de novo phospholipid biosynthesis. Two independent LPAT2 overexpression lines showed no visible phenotype under normal conditions but showed increased root length under phosphate starvation, with no effect on phosphate starvation response including marker gene expression, root hair development and anthocyanin accumulation. Accompanying membrane glycerolipid profiling of LPAT2-overexpressing plants revealed an increased content of major phospholipid classes and distinct responses to phosphate starvation between shoot and root. The findings propose a revised model of membrane lipid remodelling, in which de novo phospholipid biosynthesis mediated by LPAT2 contributes significantly to root development under phosphate starvation. © 2017 John Wiley & Sons Ltd.

Membrane Lipid Replacement for chronic illnesses, aging and cancer using oral glycerolphospholipid formulations with fructooligosaccharides to restore phospholipid function in cellular membranes, organelles, cells and tissues.

Science.gov (United States)

Nicolson, Garth L; Ash, Michael E

2017-09-01

Membrane Lipid Replacement is the use of functional, oral supplements containing mixtures of cell membrane glycerolphospholipids, plus fructooligosaccharides (for protection against oxidative, bile acid and enzymatic damage) and antioxidants, in order to safely replace damaged, oxidized, membrane phospholipids and restore membrane, organelle, cellular and organ function. Defects in cellular and intracellular membranes are characteristic of all chronic medical conditions, including cancer, and normal processes, such as aging. Once the replacement glycerolphospholipids have been ingested, dispersed, complexed and transported, while being protected by fructooligosaccharides and several natural mechanisms, they can be inserted into cell membranes, lipoproteins, lipid globules, lipid droplets, liposomes and other carriers. They are conveyed by the lymphatics and blood circulation to cellular sites where they are endocytosed or incorporated into or transported by cell membranes. Inside cells the glycerolphospholipids can be transferred to various intracellular membranes by lipid globules, liposomes, membrane-membrane contact or by lipid carrier transfer. Eventually they arrive at their membrane destinations due to 'bulk flow' principles, and there they can stimulate the natural removal and replacement of damaged membrane lipids while undergoing further enzymatic alterations. Clinical trials have shown the benefits of Membrane Lipid Replacement in restoring mitochondrial function and reducing fatigue in aged subjects and chronically ill patients. Recently Membrane Lipid Replacement has been used to reduce pain and other symptoms as well as removing hydrophobic chemical contaminants, suggesting that there are additional new uses for this safe, natural medicine supplement. This article is part of a Special Issue entitled: Membrane Lipid Therapy: Drugs Targeting Biomembranes edited by Pablo V. Escribá. Copyright © 2017 The Author(s). Published by Elsevier B.V. All rights

Suppression of phospholipid biosynthesis by cerulenin in the condensed Single-Protein-Production (cSPP) system

International Nuclear Information System (INIS)

Mao, Lili; Inoue, Koichi; Tao, Yisong; Montelione, Gaetano T.; McDermott, Ann E.; Inouye, Masayori

2011-01-01

Using the single-protein-production (SPP) system, a protein of interest can be exclusively produced in high yield from its ACA-less gene in Escherichia coli expressing MazF, an ACA-specific mRNA interferase. It is thus feasible to study a membrane protein by solid-state NMR (SSNMR) directly in natural membrane fractions. In developing isotope-enrichment methods, we observed that 13 C was also incorporated into phospholipids, generating spurious signals in SSNMR spectra. Notable, with the SPP system a protein can be produced in total absence of cell growth caused by antibiotics. Here, we demonstrate that cerulenin, an inhibitor of phospholipid biosynthesis, can suppress isotope incorporation in the lipids without affecting membrane protein yield in the SPP system. SSNMR analysis of ATP synthase subunit c, an E. coli inner membrane protein, produced by the SPP method using cerulenin revealed that 13 C resonance signals from phospholipid were markedly reduced, while signals for the isotope-enriched protein were clearly present.

Phospholipid composition and longevity: lessons from Ames dwarf mice.

Science.gov (United States)

Valencak, Teresa G; Ruf, Thomas

2013-12-01

Membrane fatty acid (FA) composition is correlated with longevity in mammals. The "membrane pacemaker hypothesis of ageing" proposes that animals which cellular membranes contain high amounts of polyunsaturated FAs (PUFAs) have shorter life spans because their membranes are more susceptible to peroxidation and further oxidative damage. It remains to be shown, however, that long-lived phenotypes such as the Ames dwarf mouse have membranes containing fewer PUFAs and thus being less prone to peroxidation, as would be predicted from the membrane pacemaker hypothesis of ageing. Here, we show that across four different tissues, i.e., muscle, heart, liver and brain as well as in liver mitochondria, Ames dwarf mice possess membrane phospholipids containing between 30 and 60 % PUFAs (depending on the tissue), which is similar to PUFA contents of their normal-sized, short-lived siblings. However, we found that that Ames dwarf mice membrane phospholipids were significantly poorer in n-3 PUFAs. While lack of a difference in PUFA contents is contradicting the membrane pacemaker hypothesis, the lower n-3 PUFAs content in the long-lived mice provides some support for the membrane pacemaker hypothesis of ageing, as n-3 PUFAs comprise those FAs being blamed most for causing oxidative damage. By comparing tissue composition between 1-, 2- and 6-month-old mice in both phenotypes, we found that membranes differed both in quantity of PUFAs and in the prevalence of certain PUFAs. In sum, membrane composition in the Ames dwarf mouse supports the concept that tissue FA composition is related to longevity.

Softening of phospholipid membranes by the adhesion of silica nanoparticles - as seen by neutron spin-echo (NSE)

Science.gov (United States)

Hoffmann, Ingo; Michel, Raphael; Sharp, Melissa; Holderer, Olaf; Appavou, Marie-Sousai; Polzer, Frank; Farago, Bela; Gradzielski, Michael

2014-05-01

The interactions between nanoparticles and vesicles are of significant interest both from a fundamental as well as from a practical point of view, as vesicles can serve as a model system for cell membranes. Accordingly the effect of nanoparticles that bind to the vesicle bilayer is very important with respect to understanding their biological impact and also may shed some light on the mechanisms behind the effect of nanotoxicity. In this study we have investigated the influence of small adsorbed silica nanoparticles (SiNPs) on the structure of zwitterionic DOPC vesicles. By a combination of SANS, cryo-TEM, and DLS, we observed that the SiNPs are bound to the outer vesicle surface without significantly affecting the vesicle structure. Most interestingly, by means of neutron spin-echo (NSE) local bilayer fluctuations were studied and one finds a small but marked decrease of the membrane rigidity upon binding of the nanoparticles. This surprising finding may be a relevant aspect for the further understanding of the effects that nanoparticles have on phospholipid bilayers.The interactions between nanoparticles and vesicles are of significant interest both from a fundamental as well as from a practical point of view, as vesicles can serve as a model system for cell membranes. Accordingly the effect of nanoparticles that bind to the vesicle bilayer is very important with respect to understanding their biological impact and also may shed some light on the mechanisms behind the effect of nanotoxicity. In this study we have investigated the influence of small adsorbed silica nanoparticles (SiNPs) on the structure of zwitterionic DOPC vesicles. By a combination of SANS, cryo-TEM, and DLS, we observed that the SiNPs are bound to the outer vesicle surface without significantly affecting the vesicle structure. Most interestingly, by means of neutron spin-echo (NSE) local bilayer fluctuations were studied and one finds a small but marked decrease of the membrane rigidity upon

Hybrid, Nanoscale Phospholipid/Block Copolymer Vesicles

Directory of Open Access Journals (Sweden)

Bo Liedberg

2013-09-01

Full Text Available Hybrid phospholipid/block copolymer vesicles, in which the polymeric membrane is blended with phospholipids, display interesting self-assembly behavior, incorporating the robustness and chemical versatility of polymersomes with the softness and biocompatibility of liposomes. Such structures can be conveniently characterized by preparing giant unilamellar vesicles (GUVs via electroformation. Here, we are interested in exploring the self-assembly and properties of the analogous nanoscale hybrid vesicles (ca. 100 nm in diameter of the same composition prepared by film-hydration and extrusion. We show that the self-assembly and content-release behavior of nanoscale polybutadiene-b-poly(ethylene oxide (PB-PEO/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC hybrid phospholipid/block copolymer vesicles can be tuned by the mixing ratio of the amphiphiles. In brief, these hybrids may provide alternative tools for drug delivery purposes and molecular imaging/sensing applications and clearly open up new avenues for further investigation.

Slaved diffusion in phospholipid bilayers

Science.gov (United States)

Zhang, Liangfang; Granick, Steve

2005-01-01

The translational diffusion of phospholipids in supported fluid bilayers splits into two populations when polyelectrolytes adsorb at incomplete surface coverage. Spatially resolved measurements using fluorescence correlation spectroscopy show that a slow mode, whose magnitude scales inversely with the degree of polymerization of the adsorbate, coexists with a fast mode characteristic of naked lipid diffusion. Inner and outer leaflets of the bilayer are affected nearly equally. Mobility may vary from spot to spot on the membrane surface, despite the lipid composition being the same. This work offers a mechanism to explain how nanosized domains with reduced mobility arise in lipid membranes. PMID:15967988

The herpes simplex virus 1 U{sub S}3 regulates phospholipid synthesis

Energy Technology Data Exchange (ETDEWEB)

Wild, Peter, E-mail: pewild@access.uzh.ch [Institute of Veterinary Anatomy, University of Zuerich (Switzerland); Institute of Virology, University of Zuerich (Switzerland); Oliveira, Anna Paula de [Institute of Virology, University of Zuerich (Switzerland); Sonda, Sabrina [Institute for Parasitology, University of Zuerich (Switzerland); Schraner, Elisabeth M. [Institute of Veterinary Anatomy, University of Zuerich (Switzerland); Institute of Virology, University of Zuerich (Switzerland); Ackermann, Mathias; Tobler, Kurt [Institute of Virology, University of Zuerich (Switzerland)

2012-10-25

Herpes simplex virus type 1 capsids bud at nuclear and Golgi membranes for envelopment by phospholipid bilayers. In the absence of U{sub S}3, nuclear membranes form multiple folds harboring virions that suggests disturbance in membrane turnover. Therefore, we investigated phospholipid metabolism in cells infected with the U{sub S}3 deletion mutant R7041({Delta}U{sub S}3), and quantified membranes involved in viral envelopment. We report that (i) [{sup 3}H]-choline incorporation into nuclear membranes and cytoplasmic membranes was enhanced peaking at 12 or 20 h post inoculation with wild type HSV-1 and R7041({Delta}U{sub S}3), respectively, (ii) the surface area of nuclear membranes increased until 24 h of R7041({Delta}U{sub S}3) infection forming folds that equaled {approx}45% of the nuclear surface, (iii) the surface area of viral envelopes between nuclear membranes equaled {approx}2400 R7041({Delta}U{sub S}3) virions per cell, and (iv) during R7041({Delta}U{sub S}3) infection, the Golgi complex expanded dramatically. The data indicate that U{sub S}3 plays a significant role in regulation of membrane biosynthesis.

Phospholipid composition of cell-derived microparticles determined by one-dimensional high-performance thin-layer chromatography

NARCIS (Netherlands)

Weerheim, A. M.; Kolb, A. M.; Sturk, A.; Nieuwland, R.

2002-01-01

Microparticles in the circulation activate the coagulation system and may activate the complement system via C-reactive protein upon conversion of membrane phospholipids by phospholipases. We developed a sensitive and reproducible method to determine the phospholipid composition of microparticles.

Binding of Diphtheria Toxin to Phospholipids in Liposomes

Science.gov (United States)

Alving, Carl R.; Iglewski, Barbara H.; Urban, Katharine A.; Moss, Joel; Richards, Roberta L.; Sadoff, Jerald C.

1980-04-01

Diphtheria toxin bound to the phosphate portion of some, but not all, phospholipids in liposomes. Liposomes consisting of dimyristoyl phosphatidylcholine and cholesterol did not bind toxin. Addition of 20 mol% (compared to dimyristoyl phosphatidylcholine) of dipalmitoyl phosphatidic acid, dicetyl phosphate, phosphatidylinositol phosphate, cardiolipin, or phosphatidylserine in the liposomes resulted in substantial binding of toxin. Inclusion of phosphatidylinositol in dimyristol phosphatidylcholine / cholesterol liposomes did not result in toxin binding. The calcium salt of dipalmitoyl phosphatidic acid was more effective than the sodium salt, and the highest level of binding occurred with liposomes consisting only of dipalmitoyl phosphatidic acid (calcium salt) and cholesterol. Binding of toxin to liposomes was dependent on pH, and the pattern of pH dependence varied with liposomes having different compositions. Incubation of diphtheria toxin with liposomes containing dicetyl phosphate resulted in maximal binding at pH 3.6, whereas binding to liposomes containing phosphatidylinositol phosphate was maximal above pH 7. Toxin did not bind to liposomes containing 20 mol% of a free fatty acid (palmitic acid) or a sulfated lipid (3-sulfogalactosylceramide). Toxin binding to dicetyl phosphate or phosphatidylinositol phosphate was inhibited by UTP, ATP, phosphocholine, or p-nitrophenyl phosphate, but not by uracil. We conclude that (a) diphtheria toxin binds specifically to the phosphate portion of certain phospholipids, (b) binding to phospholipids in liposomes is dependent on pH, but is not due only to electrostatic interaction, and (c) binding may be strongly influenced by the composition of adjacent phospholipids that do not bind toxin. We propose that a minor membrane phospholipid (such as phosphatidylinositol phosphate or phosphatidic acid), or that some other phosphorylated membrane molecule (such as a phosphoprotein) may be important in the initial binding of

Evolution of phospholipid contents during the production of quark cheese from buttermilk.

Science.gov (United States)

Ferreiro, T; Martínez, S; Gayoso, L; Rodríguez-Otero, J L

2016-06-01

We report the evolution of phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylcholine (PC), phosphatidylserine (PS), and sphingomyelin (SM) contents during the production of quark cheese from buttermilk by successive ultrafiltration concentration, enrichment with cream, concurrent homogenization and pasteurization, fermentative coagulation, and separation of quark from whey by further ultrafiltration. Buttermilk is richer than milk itself in phospholipids that afford desirable functional and technological properties, and is widely used in dairy products. To investigate how phospholipid content is affected by end-product production processes such as ultrafiltration, homogenization, pasteurization or coagulation, we measured the phospholipids at several stages of each of 5 industrial-scale quark cheese production runs. In each run, 10,000L of buttermilk was concentrated to half volume by ultrafiltration, enriched with cream, homogenized, pasteurized, inoculated with lactic acid bacteria, incubated to coagulation, and once more concentrated to half volume by ultrafiltration. Phospholipid contents were determined by HPLC with evaporative light scattering detection in the starting buttermilk, concentrated buttermilk, ultrafiltrate, cream-enriched concentrated buttermilk (both before and after concurrent homogenization and pasteurization), coagulate, and quark, and also in the rinsings obtained when the ultrafiltration equipment was washed following initial concentration. The average phospholipid content of buttermilk was approximately 5 times that of milk, and the phospholipid content of buttermilk fat 26 to 29 times that of milk fat. Although phospholipids did not cross ultrafiltration membranes, significant losses occurred during ultrafiltration (due to retention on the membranes) and during the homogenization and pasteurization process. During coagulation, however, phospholipid content rose, presumably as a consequence of the proliferation of the

Monte Carlo simulations of the distributions of intra- and extra-vesicular ions and membrane associated charges in hybrid liposomes composed of negatively charged tetraether and zwitterionic diester phospholipids

Directory of Open Access Journals (Sweden)

István P. Sugár

2017-04-01

Full Text Available Here, we model a negatively charged lipid vesicle, composed of a mixture of bipolar tetraether and diester (or diether phospholipid molecules, by a spherical shell that has zero ion permeability. We take into consideration all the charge-charge interactions between intra-vesicular ions, extra-vesicular ions, and membrane lipid associated charges. Monte Carlo simulations result in homogeneous and double-exponential ion distribution, respectively, in the intra- and extra-vesicular space. The extra-vesicular ion concentration close to the membrane surface is proportional to the total amount of the membrane charges (Nm and is independent of the partitioning of the membrane charges between the outer (Nom and inner membrane (Nim surface. This result shows that one should not disregard the effect of the charges on the inner membrane surface when calculating the ion distributions around a charged vesicle. If the partitioning of the membrane charges is not restricted (i.e., lipid flip-flop is allowed, then at different Nm, the Nom/Nim ratio remains constant and the value of Nom/Nim, as a consequence of the interaction between every charges of the model, is close to, but significantly higher than, the ratio of the outer to the inner surface area of the membrane. These results indicate that the amount and the orientation of the negatively-charged tetraether lipids in the membrane are important determinants of membrane properties in tetraether/zwitterionic diester phospholipid liposomes. Finally we compared the results of our discrete charge model and continuous models based on the solutions of the Poisson-Boltzmann equation and pointed out qualitative similarities and sometimes major quantitative differences between these two types of models.

Finite element method (FEM) model of the mechanical stress on phospholipid membranes from shock waves produced in nanosecond electric pulses (nsEP)

Science.gov (United States)

Barnes, Ronald; Roth, Caleb C.; Shadaram, Mehdi; Beier, Hope; Ibey, Bennett L.

2015-03-01

The underlying mechanism(s) responsible for nanoporation of phospholipid membranes by nanosecond pulsed electric fields (nsEP) remains unknown. The passage of a high electric field through a conductive medium creates two primary contributing factors that may induce poration: the electric field interaction at the membrane and the shockwave produced from electrostriction of a polar submersion medium exposed to an electric field. Previous work has focused on the electric field interaction at the cell membrane, through such models as the transport lattice method. Our objective is to model the shock wave cell membrane interaction induced from the density perturbation formed at the rising edge of a high voltage pulse in a polar liquid resulting in a shock wave propagating away from the electrode toward the cell membrane. Utilizing previous data from cell membrane mechanical parameters, and nsEP generated shockwave parameters, an acoustic shock wave model based on the Helmholtz equation for sound pressure was developed and coupled to a cell membrane model with finite-element modeling in COMSOL. The acoustic structure interaction model was developed to illustrate the harmonic membrane displacements and stresses resulting from shockwave and membrane interaction based on Hooke's law. Poration is predicted by utilizing membrane mechanical breakdown parameters including cortical stress limits and hydrostatic pressure gradients.

Impact of the β-Lactam Resistance Modifier (−-Epicatechin Gallate on the Non-Random Distribution of Phospholipids across the Cytoplasmic Membrane of Staphylococcus aureus

Directory of Open Access Journals (Sweden)

Helena Rosado

2015-07-01

Full Text Available The polyphenol (−-epicatechin gallate (ECg inserts into the cytoplasmic membrane (CM of methicillin-resistant Staphylococcus aureus (MRSA and reversibly abrogates resistance to β-lactam antibiotics. ECg elicits an increase in MRSA cell size and induces thickened cell walls. As ECg partially delocalizes penicillin-binding protein PBP2 from the septal division site, reduces PBP2 and PBP2a complexation and induces CM remodelling, we examined the impact of ECg membrane intercalation on phospholipid distribution across the CM and determined if ECg affects the equatorial, orthogonal mode of division. The major phospholipids of the staphylococcal CM, lysylphosphatidylglycerol (LPG, phosphatidylglycerol (PG, and cardiolipin (CL, were distributed in highly asymmetric fashion; 95%–97% of LPG was associated with the inner leaflet whereas PG (~90% and CL (~80% were found predominantly in the outer leaflet. ECg elicited small, significant changes in LPG distribution. Atomic force microscopy established that ECg-exposed cells divided in similar fashion to control bacteria, with a thickened band of encircling peptidoglycan representing the most recent plane of cell division, less distinct ribs indicative of previous sites of orthogonal division and concentric rings and “knobbles” representing stages of peptidoglycan remodelling during the cell cycle. Preservation of staphylococcal membrane lipid asymmetry and mode of division in sequential orthogonal planes appear key features of ECg-induced stress.

Effect of polyunsaturated fatty acids and phospholipids on [3H]-vitamin E incorporation into pulmonary artery endothelial cell membranes

International Nuclear Information System (INIS)

Sekharam, K.M.; Patel, J.M.; Block, E.R.

1990-01-01

Vitamin E, a dietary antioxidant, is presumed to be incorporated into the lipid bilayer of biological membranes to an extent proportional to the amount of polyunsaturated fatty acids or phospholipids in the membrane. In the present study we evaluated the distribution of incorporated polyunsaturated fatty acids (PUFA) and phosphatidylethanolamine (PE) in various membranes of pulmonary artery endothelial cells. We also studied whether incorporation of PUFA or PE is responsible for increased incorporation of [3H]-vitamin E into the membranes of these cells. Following a 24-hr incubation with linoleic acid (18:2), 18:2 was increased by 6.9-, 9.2-, and 13.2-fold in plasma, mitochondrial, and microsomal membranes, respectively. Incorporation of 18:2 caused significant increases in the unsaturation indexes of mitochondrial and microsomal polyunsaturated fatty acyl chains (P less than .01 versus control in both membranes). Incubation with arachidonic acid (20:4) for 24 hr resulted in 1.5-, 2.3-, and 2.4-fold increases in 20:4 in plasma, mitochondrial, and microsomal membranes, respectively. The unsaturation indexes of polyunsaturated fatty acyl chains of mitochondrial and microsomal membranes also increased (P less than .01 versus control in both membranes). Although incubations with 18:2 or 20:4 resulted in several-fold increases in membrane 18:2 or 20:4 fatty acids, incorporation of [3H]-vitamin E into these membranes was similar to that in controls. Following a 24-hr incubation with PE, membrane PE content was significantly increased, and [3H]-vitamin E incorporation was also increased to a comparable degree, i.e., plasma membrane greater than mitochondria greater than microsomes. Endogenous vitamin E content of the cells was not altered because of increased incorporation of PE and [3H]-vitamin E

Analysis of the induction of the myelin basic protein binding to the plasma membrane phospholipid monolayer

International Nuclear Information System (INIS)

Zhang Lei; Hao Changchun; Feng Ying; Gao Feng; Lu Xiaolong; Li Junhua; Sun Runguang

2016-01-01

Myelin basic protein (MBP) is an essential structure involved in the generation of central nervous system (CNS) myelin. Myelin shape has been described as liquid crystal structure of biological membrane. The interactions of MBP with monolayers of different lipid compositions are responsible for the multi-lamellar structure and stability of myelin. In this paper, we have designed MBP-incorporated model lipid monolayers and studied the phase behavior of MBP adsorbed on the plasma membrane at the air/water interface by thermodynamic method and atomic force microscopy (AFM). By analyzing the pressure–area ( π – A ) and pressure–time ( π – T ) isotherms, univariate linear regression equation was obtained. In addition, the elastic modulus, surface pressure increase, maximal insertion pressure, and synergy factor of monolayers were detected. These parameters can be used to modulate the monolayers binding of protein, and the results show that MBP has the strongest affinity for 1,2-dipalmitoyl-sn-glycero-3- phosphoserine (DPPS) monolayer, followed by DPPC/DPPS mixed and 1,2-dipalmitoyl-sn-glycero-3-phospho-choline (DPPC) monolayers via electrostatic and hydrophobic interactions. AFM images of DPPS and DPPC/DPPS mixed monolayers in the presence of MBP (5 nM) show a phase separation texture at the surface pressure of 20 mN/m and the incorporation of MBP put into the DPPC monolayers has exerted a significant effect on the domain structure. MBP is not an integral membrane protein but, due to its positive charge, interacts with the lipid head groups and stabilizes the membranes. The interaction between MBP and phospholipid membrane to determine the nervous system of the disease has a good biophysical significance and medical value. (special topic)

Electrochemistry of acid-base reactions in anhydrous hydrogen fluoride

International Nuclear Information System (INIS)

Masson, J.P.; Devynck, J.; Tremillon, B.

1975-01-01

Electrochemical studies were made in following media: water-HF mixtures, anhydrous HF and KF solutions in HF, solutions of the SbF5 type in HF. The acidity level of these solutions was evaluated using the R(H) functions based on the strehlow hypotheses. From the pH measurement in anhydrous HF, it was possible to get acid-base titration curves and pH buffers. The behavior of quinones in anhydrous HF is presented [fr

The γ-irradiation stability of anhydrous ethylenediamine

International Nuclear Information System (INIS)

Zhong Zhijing; Long Suqun; Fu Yibei; Xu Yunshu

2005-01-01

Anhydrous Ethylenediamine (AEDA) was irradiated in air or nitrogen at ambient temperatures by 60 Co γ-rays. The radiation degradation products were analyzed by means of GC, GC/MS, MS and FT-IR, etc. Notable gaseous products such as hydrogen, methane, ethylene, ammonia, and some volatile liquid state radiolysis products as well, were examined, and many kinds of polymerized products were detected in the liquid remain sample. It was found that under certain conditions both the hydrogen and methane contents of the products increased with dose. Remarkable effects of radiation degradation and radiation polymerization were observed, and even severe effects were found in nitrogen ambience. Therefore, a worse γ-irradiation stability of anhydrous ethylenediamine was demonstrated. The radiation polymerization of anhydrous ethylenediamine was not reported before. (authors)

Ice formation in model biological membranes in the presence of cryoprotectors

Energy Technology Data Exchange (ETDEWEB)

Kiselev, M.A. E-mail: kiselev@nf.jinr.ru; Lesieur, P.; Kisselev, A.M.; Ollivon, M

2000-06-21

Ice formation in model biological membranes is studied by SAXS and WAXS in the presence of cryoprotectors: dimethyl sulfoxide and glycerol. Three types of phospholipid membranes: DPPC, DMPC, DSPC are chosen for the investigation as well-studied model biological membranes. A special cryostat is used for sample cooling from 14.1 deg. C to -55.4 deg. C. The ice formation is detected only by WAXS in binary phospholipid/water and ternary phospholipid/cryoprotector/water systems in the condition of excess solvent. Ice formation in a binary phospholipid/water system creates an abrupt decrease of the membrane repeat distance by {delta}d, the so-called ice-induced dehydration of intermembrane space. The value of {delta}d decreases as the cryoprotector concentration increases. The formation of ice does not influence the membrane structure ({delta}d=0) for cryoprotector mole fractions higher than 0.05.

Phospholipid Complex Technique for Superior Bioavailability of Phytoconstituents

Directory of Open Access Journals (Sweden)

Kattamanchi Gnananath

2017-04-01

Full Text Available Phytoconstituents have been utilized as medicines for thousands of years, yet their application is limited owing to major hurdles like deficit lipid solubility, large molecular size and degradation in the gastric environment of gut. Recently, phospholipid-complex technique has unveiled in addressing these stumbling blocks either by enhancing the solubilizing capacity or its potentiating ability to pass through the biological membranes and it also protects the active herbal components from degradation. Hence, this phospholipid-complex-technique can enable researchers to deliver the phytoconstituents into systemic circulation by using certain conventional dosage forms like tablets and capsules. This review highlights the unique property of phospholipids in drug delivery, their role as adjuvant in health benefits, and their application in the herbal medicine systems to improve the bioavailability of active herbal components. Also we summarize the prerequisites for phytosomes preparation like the selection of type of phytoconstituents, solvents used, various methods employed in phytosomal preparation and its characterization. Further we discuss the key findings of recent research work conducted on phospholipid-based delivery systems which can enable new directions and advancements to the development of herbal dosage forms.

An averaged polarizable potential for multiscale modeling in phospholipid membranes

DEFF Research Database (Denmark)

Witzke, Sarah; List, Nanna Holmgaard; Olsen, Jógvan Magnus Haugaard

2017-01-01

A set of average atom-centered charges and polarizabilities has been developed for three types of phospholipids for use in polarizable embedding calculations. The lipids investigated are 1,2-dimyristoyl-sn-glycero-3-phosphocholine, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, and 1-palmitoyl...

Novel composite membranes based on PBI and dicationic ionic liquids for high temperature polymer electrolyte membrane fuel cells

International Nuclear Information System (INIS)

Hooshyari, Khadijeh; Javanbakht, Mehran; Adibi, Mina

2016-01-01

Two types of innovative composite membranes based on polybenzimidazole (PBI) containing dicationic ionic liquid 1,3-di(3-methylimidazolium) propane bis (trifluoromethylsulfonyl) imide (PDC 3 ) and monocationic ionic liquid 1-hexyl-3-methylimidazolium bis (trifluoromethanesulfonyl) imide (PMC 6 ) are prepared as electrolyte for high temperature fuel cells applications under anhydrous conditions. The analyses of results display promising characteristics such as high proton conductivity and thermal stability. Moreover the fuel cell performance of PA doped PDC 3 composite membranes is enhanced in comparison with PA doped PMC 6 and PA doped PBI membranes at high temperatures. Dicationic ionic liquid with high number of charge carriers provides well-developed ionic channels which form facile pathways and considerably develop the anhydrous proton conductivity. The highest proton conductivity of 81 mS/cm is achieved for PA doped PDC 3 composite membranes with PBI/IL mole ratio: 4 at 180 °C. A power density of 0.44 W/cm 2 is obtained at 0.5 V and 180 °C for PA doped PDC 3 composite membranes, which proves that these developed composite membranes can be considered as most promising candidates for high temperature fuel cell applications with enhanced proton conductivity.

49 CFR 173.195 - Hydrogen cyanide, anhydrous, stabilized (hydrocyanic acid, aqueous solution).

Science.gov (United States)

2010-10-01

... 49 Transportation 2 2010-10-01 2010-10-01 false Hydrogen cyanide, anhydrous, stabilized... Hazardous Materials Other Than Class 1 and Class 7 § 173.195 Hydrogen cyanide, anhydrous, stabilized (hydrocyanic acid, aqueous solution). (a) Hydrogen cyanide, anhydrous, stabilized, must be packed in...

Phospholipid-binding protein EhC2A mediates calcium-dependent translocation of transcription factor URE3-BP to the plasma membrane of Entamoeba histolytica.

Science.gov (United States)

Moreno, Heriberto; Linford, Alicia S; Gilchrist, Carol A; Petri, William A

2010-05-01

The Entamoeba histolytica upstream regulatory element 3-binding protein (URE3-BP) is a transcription factor that binds DNA in a Ca(2+)-inhibitable manner. The protein is located in both the nucleus and the cytoplasm but has also been found to be enriched in the plasma membrane of amebic trophozoites. We investigated the reason for the unusual localization of URE3-BP at the amebic plasma membrane. Here we identify and characterize a 22-kDa Ca(2+)-dependent binding partner of URE3-BP, EhC2A, a novel member of the C2-domain superfamily. Immunoprecipitations of URE3-BP and EhC2A showed that the proteins interact and that such interaction was enhanced in the presence of Ca(2+). Recombinant and native EhC2A bound phospholipid liposomes in a Ca(2+)-dependent manner, with half-maximal binding occurring at 3.4 muM free Ca(2+). A direct interaction between EhC2A and URE3-BP was demonstrated by the ability of recombinant EhC2A to recruit recombinant URE3-BP to phospholipid liposomes in a Ca(2+)-dependent manner. URE3-BP and EhC2A were observed to translocate to the amebic plasma membrane upon an increase in the intracellular Ca(2+) concentration of trophozoites, as revealed by subcellular fractionation and immunofluorescent staining. Short hairpin RNA-mediated knockdown of EhC2A protein expression significantly modulated the mRNA levels of URE3-BP-regulated transcripts. Based on these results, we propose a model for EhC2A-mediated regulation of the transcriptional activities of URE3-BP via Ca(2+)-dependent anchoring of the transcription factor to the amebic plasma membrane.

Frequency-dependent electrodeformation of giant phospholipid vesicles in AC electric field

Science.gov (United States)

2010-01-01

A model of vesicle electrodeformation is described which obtains a parametrized vesicle shape by minimizing the sum of the membrane bending energy and the energy due to the electric field. Both the vesicle membrane and the aqueous media inside and outside the vesicle are treated as leaky dielectrics, and the vesicle itself is modeled as a nearly spherical shape enclosed within a thin membrane. It is demonstrated (a) that the model achieves a good quantitative agreement with the experimentally determined prolate-to-oblate transition frequencies in the kilohertz range and (b) that the model can explain a phase diagram of shapes of giant phospholipid vesicles with respect to two parameters: the frequency of the applied alternating current electric field and the ratio of the electrical conductivities of the aqueous media inside and outside the vesicle, explored in a recent paper (S. Aranda et al., Biophys J 95:L19–L21, 2008). A possible use of the frequency-dependent shape transitions of phospholipid vesicles in conductometry of microliter samples is discussed. PMID:21886342

Transport of anhydrous ammoniac - risk assessment

International Nuclear Information System (INIS)

Castellano, S.; Degrange, J.P.; Hubert, P.; Pages, P.; Lamblin, J.

1989-12-01

This risk analysis of anhydrous ammonia transport in France was done within a study initiated by the Department of dangerous goods of the Ministry of transport. The study deals with the road and rail transportation of bulk anhydrous ammonia. After analysis of transport system and traffic, the transport accident risks are estimated, as well as their distribution on the French territory. Finally after a synthesis of results, a number of safety measures to be undertaken were identified. This is a joint study of SMC-CEPN, with a specific role of SEMA-METRA-CONSEIL concerning the traffic frequency, and the center for risk evaluation concerning nuclear safety

Characterisation of the membrane affinity of an isoniazide peptide conjugate by tensiometry, atomic force microscopy and sum-frequency vibrational spectroscopy, using a phospholipid Langmuir monolayer model.

Science.gov (United States)

Hill, Katalin; Pénzes, Csanád Botond; Schnöller, Donát; Horváti, Kata; Bosze, Szilvia; Hudecz, Ferenc; Keszthelyi, Tamás; Kiss, Eva

2010-10-07

Tensiometry, sum-frequency vibrational spectroscopy, and atomic force microscopy were employed to assess the cell penetration ability of a peptide conjugate of the antituberculotic agent isoniazide. Isoniazide was conjugated to peptide (91)SEFAYGSFVRTVSLPV(106), a functional T-cell epitope of the immunodominant 16 kDa protein of Mycobacterium tuberculosis. As a simple but versatile model of the cell membrane a phospholipid Langmuir monolayer at the liquid/air interface was used. Changes induced in the structure of the phospholipid monolayer by injection of the peptide conjugate into the subphase were followed by tensiometry and sum-frequency vibrational spectroscopy. The drug penetrated lipid films were transferred to a solid support by the Langmuir-Blodgett technique, and their structures were characterized by atomic force microscopy. Peptide conjugation was found to strongly enhance the cell penetration ability of isoniazide.

Protein-membrane interaction and fatty acid transfer from intestinal fatty acid-binding protein to membranes. Support for a multistep process.

Science.gov (United States)

Falomir-Lockhart, Lisandro J; Laborde, Lisandro; Kahn, Peter C; Storch, Judith; Córsico, Betina

2006-05-19

Fatty acid transfer from intestinal fatty acid-binding protein (IFABP) to phospholipid membranes occurs during protein-membrane collisions. Electrostatic interactions involving the alpha-helical "portal" region of the protein have been shown to be of great importance. In the present study, the role of specific lysine residues in the alpha-helical region of IFABP was directly examined. A series of point mutants in rat IFABP was engineered in which the lysine positive charges in this domain were eliminated or reversed. Using a fluorescence resonance energy transfer assay, we analyzed the rates and mechanism of fatty acid transfer from wild type and mutant proteins to acceptor membranes. Most of the alpha-helical domain mutants showed slower absolute fatty acid transfer rates to zwitterionic membranes, with substitution of one of the lysines of the alpha2 helix, Lys27, resulting in a particularly dramatic decrease in the fatty acid transfer rate. Sensitivity to negatively charged phospholipid membranes was also reduced, with charge reversal mutants in the alpha2 helix the most affected. The results support the hypothesis that the portal region undergoes a conformational change during protein-membrane interaction, which leads to release of the bound fatty acid to the membrane and that the alpha2 segment is of particular importance in the establishment of charge-charge interactions between IFABP and membranes. Cross-linking experiments with a phospholipid-photoactivable reagent underscored the importance of charge-charge interactions, showing that the physical interaction between wild-type intestinal fatty acid-binding protein and phospholipid membranes is enhanced by electrostatic interactions. Protein-membrane interactions were also found to be enhanced by the presence of ligand, suggesting different collisional complex structures for holo- and apo-IFABP.

INVESTIGATION ON THE MORPHOLOGY AND PROPERTIES OF AGGREGATE STRUCTURES OF NATURAL PHOSPHOLIPIDS IN AQUEOUS SYSTEM USING CRYO-TEM

Directory of Open Access Journals (Sweden)

Dwi Hudiyanti

2012-02-01

Full Text Available Cryogenic transmission electron microscopy (Cryo-TEM was used to investigate the aggregates morphology and properties of candle tree (Aleurites moluccana endosperm, sesame (Sesamum indicum L. syn. seeds, and coconut (Cocos nucifera endosperm phospholipids in dilute aqueous system. The micrographs showed that candle tree phospholipids formed planar bilayer and cluster of vesicles with lipid droplets, while coconut and sesame phospholipids formed well-defined unilamellar vesicles. The vesicles size could be as small as 50 nm in diameter. Coconut phospholipids also showed a good bending ability. Formation of clusters of vesicles was also found in coconut phospholipids dispersion, but this cluster was easily broken by extrusion through a small pore membrane.

Analysis of the induction of the myelin basic protein binding to the plasma membrane phospholipid monolayer

Science.gov (United States)

Zhang, Lei; Hao, Changchun; Feng, Ying; Gao, Feng; Lu, Xiaolong; Li, Junhua; Sun, Runguang

2016-09-01

Myelin basic protein (MBP) is an essential structure involved in the generation of central nervous system (CNS) myelin. Myelin shape has been described as liquid crystal structure of biological membrane. The interactions of MBP with monolayers of different lipid compositions are responsible for the multi-lamellar structure and stability of myelin. In this paper, we have designed MBP-incorporated model lipid monolayers and studied the phase behavior of MBP adsorbed on the plasma membrane at the air/water interface by thermodynamic method and atomic force microscopy (AFM). By analyzing the pressure-area (π-A) and pressure-time (π-T) isotherms, univariate linear regression equation was obtained. In addition, the elastic modulus, surface pressure increase, maximal insertion pressure, and synergy factor of monolayers were detected. These parameters can be used to modulate the monolayers binding of protein, and the results show that MBP has the strongest affinity for 1,2-dipalmitoyl-sn-glycero-3- phosphoserine (DPPS) monolayer, followed by DPPC/DPPS mixed and 1,2-dipalmitoyl-sn-glycero-3-phospho-choline (DPPC) monolayers via electrostatic and hydrophobic interactions. AFM images of DPPS and DPPC/DPPS mixed monolayers in the presence of MBP (5 nM) show a phase separation texture at the surface pressure of 20 mN/m and the incorporation of MBP put into the DPPC monolayers has exerted a significant effect on the domain structure. MBP is not an integral membrane protein but, due to its positive charge, interacts with the lipid head groups and stabilizes the membranes. The interaction between MBP and phospholipid membrane to determine the nervous system of the disease has a good biophysical significance and medical value. Project supported by the National Natural Science Foundation of China (Grant Nos. 21402114 and 11544009), the Natural Science Basic Research Plan in Shaanxi Province of China (Grant No. 2016JM2010), the Fundamental Research Funds for the Central

Phosphatidic acid is a major phospholipid class in reproductive organs of Arabidopsis thaliana.

Science.gov (United States)

Yunus, Ian Sofian; Cazenave-Gassiot, Amaury; Liu, Yu-Chi; Lin, Ying-Chen; Wenk, Markus R; Nakamura, Yuki

2015-01-01

Phospholipids are the crucial components of biological membranes and signal transduction. Among different tissues, flower phospholipids are one of the least characterized features of plant lipidome. Here, we report that floral reproductive organs of Arabidopsis thaliana contain high levels of phosphatidic acid (PA), a known lipid second messenger. By using floral homeotic mutants enriched with specific floral organs, lipidomics study showed increased levels of PA species in ap3-3 mutant with enriched pistils. Accompanied gene expression study for 7 diacylglycerol kinases and 11 PA phosphatases revealed distinct floral organ specificity, suggesting an active phosphorylation/dephosphorylation between PA and diacylglycerol in flowers. Our results suggest that PA is a major phospholipid class in floral reproductive organs of A. thaliana.

Possible mechanism of adhesion in a mica supported phospholipid bilayer

International Nuclear Information System (INIS)

Pertsin, Alexander; Grunze, Michael

2014-01-01

Phospholipid bilayers supported on hydrophilic solids like silica and mica play a substantial role in fundamental studies and technological applications of phospholipid membranes. In both cases the molecular mechanism of adhesion between the bilayer and the support is of primary interest. Since the possibilities of experimental methods in this specific area are rather limited, the methods of computer simulation acquire great importance. In this paper we use the grand canonical Monte Carlo technique and an atomistic force field to simulate the behavior of a mica supported phospholipid bilayer in pure water as a function of the distance between the bilayer and the support. The simulation reveals a possible adhesion mechanism, where the adhesion is due to individual lipid molecules that protrude from the bilayer and form widely spaced links with the support. Simultaneously, the bilayer remains separated from the bilayer by a thin water interlayer which maintains the bilayer fluidity

Phospholipid transfer activities in toad oocytes and developing embryos

International Nuclear Information System (INIS)

Rusinol, A.; Salomon, R.A.; Bloj, B.

1987-01-01

The role of lipid transfer proteins during plasma membrane biogenesis was explored. Developing amphibia embryos were used because during their growth an active plasma membrane biosynthesis occurs together with negligible mitochondrial and endoplasmic reticulum proliferation. Sonicated vesicles, containing 14 C-labeled phospholipids and 3 H-labeled triolein, as donor particles and cross-linked erythrocyte ghosts as acceptor particles were used to measure phospholipid transfer activities in unfertilized oocytes and in developing embryos of the toad Bufo arenarum. Phosphatidylcholine transfer activity in pH 5.1 supernatant of unfertilized oocytes was 8-fold higher than the activity found in female toad liver supernatant, but dropped steadily after fertilization. After 20 hr of development, at the stage of late blastula, the phosphatidylcholine transfer activity had dropped 4-fold. Unfertilized oocyte supernatant exhibited phosphatidylinositol and phosphatidylethanolamine transfer activity also, but at the late blastula stage the former had dropped 18-fold and the latter was no longer detectable under our assay conditions. Our results show that fertilization does not trigger a phospholipid transport process catalyzed by lipid transfer proteins. Moreover, they imply that 75% of the phosphatidylcholine transfer activity and more than 95% of the phosphatidylinositol and phosphatidylethanolamine transfer activities present in pH 5.1 supernatants of unfertilized oocytes may not be essential for toad embryo development. Our findings do not rule out, however, that a phosphatidylcholine-specific lipid transfer protein could be required for embryo early growth

Changes in the Fatty Acid Profile and Phospholipid Molecular Species Composition of Human Erythrocyte Membranes after Hybrid Palm and Extra Virgin Olive Oil Supplementation.

Science.gov (United States)

Pacetti, D; Gagliardi, R; Balzano, M; Frega, N G; Ojeda, M L; Borrero, M; Ruiz, A; Lucci, P

2016-07-13

This work aims to evaluate and compare, for the first time, the effects of extra virgin olive oil (EVOO) and hybrid palm oil (HPO) supplementation on the fatty acid profile and phospholipid (PL) molecular species composition of human erythrocyte membranes. Results supported the effectiveness of both HPO and EVOO supplementation (3 months, 25 mL/day) in decreasing the lipophilic index of erythrocytes with no significant differences between HPO and EVOO groups at month 3. On the other hand, the novel and rapid ultraperformance liquid chromatography-tandem mass spectrometry method used for PL analysis reveals an increase in the levels of phosphatidylcholine and phosphatidylethanolamine species esterified with polyunsaturated fatty acids. This work demonstrates the ability of both EVOO and HPO to increase the degree of unsaturation of erythrocyte membrane lipids with an improvement in membrane fluidity that could be associated with a lower risk of developing cardiovascular diseases.

Tombusviruses upregulate phospholipid biosynthesis via interaction between p33 replication protein and yeast lipid sensor proteins during virus replication in yeast

International Nuclear Information System (INIS)

Barajas, Daniel; Xu, Kai; Sharma, Monika; Wu, Cheng-Yu; Nagy, Peter D.

2014-01-01

Positive-stranded RNA viruses induce new membranous structures and promote membrane proliferation in infected cells to facilitate viral replication. In this paper, the authors show that a plant-infecting tombusvirus upregulates transcription of phospholipid biosynthesis genes, such as INO1, OPI3 and CHO1, and increases phospholipid levels in yeast model host. This is accomplished by the viral p33 replication protein, which interacts with Opi1p FFAT domain protein and Scs2p VAP protein. Opi1p and Scs2p are phospholipid sensor proteins and they repress the expression of phospholipid genes. Accordingly, deletion of OPI1 transcription repressor in yeast has a stimulatory effect on TBSV RNA accumulation and enhanced tombusvirus replicase activity in an in vitro assay. Altogether, the presented data convincingly demonstrate that de novo lipid biosynthesis is required for optimal TBSV replication. Overall, this work reveals that a (+)RNA virus reprograms the phospholipid biosynthesis pathway in a unique way to facilitate its replication in yeast cells. - Highlights: • Tombusvirus p33 replication protein interacts with FFAT-domain host protein. • Tombusvirus replication leads to upregulation of phospholipids. • Tombusvirus replication depends on de novo lipid synthesis. • Deletion of FFAT-domain host protein enhances TBSV replication. • TBSV rewires host phospholipid synthesis

Ionic protein-lipid interaction at the plasma membrane: what can the charge do?

Science.gov (United States)

Li, Lunyi; Shi, Xiaoshan; Guo, Xingdong; Li, Hua; Xu, Chenqi

2014-03-01

Phospholipids are the major components of cell membranes, but they have functional roles beyond forming lipid bilayers. In particular, acidic phospholipids form microdomains in the plasma membrane and can ionically interact with proteins via polybasic sequences, which can have functional consequences for the protein. The list of proteins regulated by ionic protein-lipid interaction has been quickly expanding, and now includes membrane proteins, cytoplasmic soluble proteins, and viral proteins. Here we review how acidic phospholipids in the plasma membrane regulate protein structure and function via ionic interactions, and how Ca(2+) regulates ionic protein-lipid interactions via direct and indirect mechanisms. Copyright © 2014 Elsevier Ltd. All rights reserved.

Nanoelectropulse-driven membrane perturbation and small molecule permeabilization

Directory of Open Access Journals (Sweden)

Sun Yinghua

2006-10-01

Full Text Available Abstract Background Nanosecond, megavolt-per-meter pulsed electric fields scramble membrane phospholipids, release intracellular calcium, and induce apoptosis. Flow cytometric and fluorescence microscopy evidence has associated phospholipid rearrangement directly with nanoelectropulse exposure and supports the hypothesis that the potential that develops across the lipid bilayer during an electric pulse drives phosphatidylserine (PS externalization. Results In this work we extend observations of cells exposed to electric pulses with 30 ns and 7 ns durations to still narrower pulse widths, and we find that even 3 ns pulses are sufficient to produce responses similar to those reported previously. We show here that in contrast to unipolar pulses, which perturb membrane phospholipid order, tracked with FM1-43 fluorescence, only at the anode side of the cell, bipolar pulses redistribute phospholipids at both the anode and cathode poles, consistent with migration of the anionic PS head group in the transmembrane field. In addition, we demonstrate that, as predicted by the membrane charging hypothesis, a train of shorter pulses requires higher fields to produce phospholipid scrambling comparable to that produced by a time-equivalent train of longer pulses (for a given applied field, 30, 4 ns pulses produce a weaker response than 4, 30 ns pulses. Finally, we show that influx of YO-PRO-1, a fluorescent dye used to detect early apoptosis and activation of the purinergic P2X7 receptor channels, is observed after exposure of Jurkat T lymphoblasts to sufficiently large numbers of pulses, suggesting that membrane poration occurs even with nanosecond pulses when the electric field is high enough. Propidium iodide entry, a traditional indicator of electroporation, occurs with even higher pulse counts. Conclusion Megavolt-per-meter electric pulses as short as 3 ns alter the structure of the plasma membrane and permeabilize the cell to small molecules. The dose

Specific Interaction between Redox Phospholipid Polymers and Plastoquinone in Photosynthetic Electron Transport Chain.

Science.gov (United States)

Tanaka, Kenya; Kaneko, Masahiro; Ishikawa, Masahito; Kato, Souichiro; Ito, Hidehiro; Kamachi, Toshiaki; Kamiya, Kazuhide; Nakanishi, Shuji

2017-04-19

Redox phospholipid polymers added in culture media are known to be capable of extracting electrons from living photosynthetic cells across bacterial cell membranes with high cytocompatibility. In the present study, we identify the intracellular redox species that transfers electrons to the polymers. The open-circuit electrochemical potential of an electrolyte containing the redox polymer and extracted thylakoid membranes shift to positive (or negative) under light irradiation, when an electron transport inhibitor specific to plastoquinone is added upstream (or downstream) in the photosynthetic electron transport chain. The same trend is also observed for a medium containing living photosynthetic cells of Synechococcus elongatus PCC7942. These results clearly indicate that the phospholipid redox polymers extract photosynthetic electrons mainly from plastoquinone. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Anisotropic amplification of proton transport in proton exchange membrane fuel cells

Science.gov (United States)

Thimmappa, Ravikumar; Fawaz, Mohammed; Devendrachari, Mruthyunjayachari Chattanahalli; Gautam, Manu; Kottaichamy, Alagar Raja; Shafi, Shahid Pottachola; Thotiyl, Musthafa Ottakam

2017-07-01

Though graphene oxide (GO) membrane shuttles protons under humid conditions, it suffer severe disintegration and anhydrous conditions lead to abysmal ionic conductivity. The trade-off between mechanical integrity and ionic conductivity challenge the amplification of GO's ionic transport under anhydrous conditions. We show anisotropic amplification of GO's ionic transport with a selective amplification of in plane contribution under anhydrous conditions by doping it with a plant extract, phytic acid (PA). The hygroscopic nature of PA stabilized interlayer water molecules and peculiar geometry of sbnd OH functionalities around saturated hydrocarbon ring anisotropically enhanced ionic transport amplifying the fuel cell performance metrics.

Phospholipid metabolism and nuclear function: roles of the lipin family of phosphatidic acid phosphatases.

Science.gov (United States)

Siniossoglou, Symeon

2013-03-01

Phospholipids play important roles in nuclear function as dynamic building blocks for the biogenesis of the nuclear membrane, as well as signals by which the nucleus communicates with other organelles, and regulate a variety of nuclear events. The mechanisms underlying the nuclear roles of phospholipids remain poorly understood. Lipins represent a family of phosphatidic acid (PA) phosphatases that are conserved from yeasts to humans and perform essential functions in lipid metabolism. Several studies have identified key roles for lipins and their regulators in nuclear envelope organization, gene expression and the maintenance of lipid homeostasis in yeast and metazoans. This review discusses recent advances in understanding the roles of lipins in nuclear structure and function. This article is part of a Special Issue entitled Phospholipids and Phospholipid Metabolism. Copyright © 2012 Elsevier B.V. All rights reserved.

Competing intermolecular interactions of artemisinin-type agents and aspirin with membrane phospholipids: Combined model mass spectrometry and quantum-chemical study

International Nuclear Information System (INIS)

Pashynska, Vlada; Stepanian, Stepan; Gömöry, Agnes; Vekey, Karoly; Adamowicz, Ludwik

2015-01-01

Highlights: • Competitive binding of artemisinin agents and aspirin with phospholipids is shown. • Complexation between the antimalarial drugs and aspirin molecules is also found. • Energetically favorable structures of the model complexes are identified by DFT. • Membranotropic activity of the studied drugs can be modified under joint usage. - Abstract: Study of intermolecular interactions of antimalarial artemisinin-type drugs and aspirin with membrane phospholipids is important in term of elucidation of the drugs activity modification under their joint usage. Combined experimental and computational study of the interaction of dihydroartemisinin, α-artemether, and artesunate with aspirin (ASP) and dipalmitoylphosphatidylcholine (DPPC) is performed by electrospray ionization (ESI) mass spectrometry and by DFT B3LYP/aug-cc-pVDZ methods. The results of the ESI investigation of systems containing artemisinin-type agent, ASP and DPPC, reveal a competition between the antimalarial agents and ASP for binding with DPPC molecules. The complexation between the antimalarial drugs and ASP is also found. Observed phenomena suggest that membranotropic activity of artemisin-type agents and aspirin is modified under their combined usage. To elucidate structure-energy characteristics of the non-covalent complexes studied the model DFT calculations are performed for dihydroartemisinin · ASP complex and complexes of the each drug with phosphatidylcholine head of DPPC in neutral and cationized forms

Competing intermolecular interactions of artemisinin-type agents and aspirin with membrane phospholipids: Combined model mass spectrometry and quantum-chemical study

Energy Technology Data Exchange (ETDEWEB)

Pashynska, Vlada, E-mail: vlada@vl.kharkov.ua [B.Verkin Institute for Low Temperature Physics and Engineering of the National Academy of Sciences of Ukraine, Lenin Ave., 47, 61103 Kharkov (Ukraine); Stepanian, Stepan [B.Verkin Institute for Low Temperature Physics and Engineering of the National Academy of Sciences of Ukraine, Lenin Ave., 47, 61103 Kharkov (Ukraine); Gömöry, Agnes; Vekey, Karoly [Institute of Organic Chemistry of Research Centre for Natural Sciences of the Hungarian Academy of Sciences, Magyar tudosok korutja, 2, Budapest H-1117 (Hungary); Adamowicz, Ludwik [University of Arizona, Department of Chemistry and Biochemistry, Tucson, AZ 85721 (United States)

2015-07-09

Highlights: • Competitive binding of artemisinin agents and aspirin with phospholipids is shown. • Complexation between the antimalarial drugs and aspirin molecules is also found. • Energetically favorable structures of the model complexes are identified by DFT. • Membranotropic activity of the studied drugs can be modified under joint usage. - Abstract: Study of intermolecular interactions of antimalarial artemisinin-type drugs and aspirin with membrane phospholipids is important in term of elucidation of the drugs activity modification under their joint usage. Combined experimental and computational study of the interaction of dihydroartemisinin, α-artemether, and artesunate with aspirin (ASP) and dipalmitoylphosphatidylcholine (DPPC) is performed by electrospray ionization (ESI) mass spectrometry and by DFT B3LYP/aug-cc-pVDZ methods. The results of the ESI investigation of systems containing artemisinin-type agent, ASP and DPPC, reveal a competition between the antimalarial agents and ASP for binding with DPPC molecules. The complexation between the antimalarial drugs and ASP is also found. Observed phenomena suggest that membranotropic activity of artemisin-type agents and aspirin is modified under their combined usage. To elucidate structure-energy characteristics of the non-covalent complexes studied the model DFT calculations are performed for dihydroartemisinin · ASP complex and complexes of the each drug with phosphatidylcholine head of DPPC in neutral and cationized forms.

Efficacy of bacterial bioremediation: Demonstration of complete incorporation of hydrocarbons into membrane phospholipids from Rhodococcus hydrocarbon degrading bacteria by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry

Energy Technology Data Exchange (ETDEWEB)

Rodgers, R.P.; Blumer, E.N.; Emmett, M.R.; Marshall, A.G.

2000-02-01

The authors present a method and example to establish complete incorporation of hydrocarbons into membrane phospholipids of putatively bioremediative bacteria. Bacteria are grown on minimal media containing a specified carbon source, either natural abundance or enriched. After extraction (but no other prior separation) of the membrane lipids, electrospray ionization yields a negative-ion FT-ICR mass spectrum containing prominent phospholipid parent ions. If {sup 13}C-enriched hydrocarbon incorporation is complete, then the mass of the parent ion will increase by n Da, in which n is the number of its constituent carbon atoms; moreover, the {sup 13}C isotopic distribution pattern will be reversed. The identities of the constituent fatty acids and polar headgroup are obtained by collisional dissociation (MS/MS), and their extent of {sup 13}C incorporation determined individually. The method is demonstrated for Rhodococcus rhodochrous (ATCC No. 53968), for which all 44 carbons of a representative phosphatidylinositol are shown to derive from the hydrocarbon source. Interestingly, although only C{sub 16} and C{sub 18} alkanes are provided in the growth medium, the bacteria synthesize uniformly enriched C16:0 and C19:0 fatty acids.

Salicylic acid induces vanillin synthesis through the phospholipid signaling pathway in Capsicum chinense cell cultures.

Science.gov (United States)

Rodas-Junco, Beatriz A; Cab-Guillén, Yahaira; Muñoz-Sánchez, J Armando; Vázquez-Flota, Felipe; Monforte-González, Miriam; Hernández-Sotomayor, S M Teresa

2013-10-01

Signal transduction via phospholipids is mediated by phospholipases such as phospholipase C (PLC) and D (PLD), which catalyze hydrolysis of plasma membrane structural phospholipids. Phospholipid signaling is also involved in plant responses to phytohormones such as salicylic acid (SA). The relationships between phospholipid signaling, SA, and secondary metabolism are not fully understood. Using a Capsicum chinense cell suspension as a model, we evaluated whether phospholipid signaling modulates SA-induced vanillin production through the activation of phenylalanine ammonia lyase (PAL), a key enzyme in the biosynthetic pathway. Salicylic acid was found to elicit PAL activity and consequently vanillin production, which was diminished or reversed upon exposure to the phosphoinositide-phospholipase C (PI-PLC) signaling inhibitors neomycin and U73122. Exposure to the phosphatidic acid inhibitor 1-butanol altered PLD activity and prevented SA-induced vanillin production. Our results suggest that PLC and PLD-generated secondary messengers may be modulating SA-induced vanillin production through the activation of key biosynthetic pathway enzymes.

Pollen viability and membrane lipid composition

NARCIS (Netherlands)

Bilsen, van D.G.J.L.

1993-01-01

In this thesis membrane lipid composition is studied in relation to pollen viability during storage. Chapter 1 reviews pollen viability, membranes in the dry state and membrane changes associated with cellular aging. This chapter is followed by a study of age-related changes in phospholipid

The Effect of Compaction Force on the Transition to Hydrate of Anhydrous Aripiprazole.

Science.gov (United States)

Togo, Taichiro; Taniguchi, Toshiya; Nakata, Yoshitaka

2018-01-01

Aripiprazole (APZ) is used to treat schizophrenia and is administered as a tablet containing the anhydrous form of APZ. In this study, the effect of compaction force on the crystal form transition was investigated. The crystalline state was observed by X-ray diffraction (XRD). APZ Anhydrous Form II was compacted into tablets. The XRD intensity of anhydrous APZ became lower with higher compressive force. The degree of crystallinity decreased with the compaction force. The powder and the compacted tablets of anhydrous APZ were stored for one week under 60°C and 75% relative humidity. The powder showed no crystal form transition after storage. For the tablets, however, XRD peaks of APZ hydrate were observed after storage. The tablets compacted with higher force showed the higher XRD diffraction intensity of hydrate form. We concluded that the crystallinity reduction of APZ Anhydrous Form II by compaction caused and accelerated the transition to hydrate under high temperature and humidity conditions. In order to manufacture crystallographically stable tablets containing anhydrous APZ, it is important to prevent this crystallinity reduction during compaction.

Layer-by-layer cell membrane assembly

Science.gov (United States)

Matosevic, Sandro; Paegel, Brian M.

2013-11-01

Eukaryotic subcellular membrane systems, such as the nuclear envelope or endoplasmic reticulum, present a rich array of architecturally and compositionally complex supramolecular targets that are as yet inaccessible. Here we describe layer-by-layer phospholipid membrane assembly on microfluidic droplets, a route to structures with defined compositional asymmetry and lamellarity. Starting with phospholipid-stabilized water-in-oil droplets trapped in a static droplet array, lipid monolayer deposition proceeds as oil/water-phase boundaries pass over the droplets. Unilamellar vesicles assembled layer-by-layer support functional insertion both of purified and of in situ expressed membrane proteins. Synthesis and chemical probing of asymmetric unilamellar and double-bilayer vesicles demonstrate the programmability of both membrane lamellarity and lipid-leaflet composition during assembly. The immobilized vesicle arrays are a pragmatic experimental platform for biophysical studies of membranes and their associated proteins, particularly complexes that assemble and function in multilamellar contexts in vivo.

Electrostatic control of phospholipid polymorphism.

Science.gov (United States)

Tarahovsky, Y S; Arsenault, A L; MacDonald, R C; McIntosh, T J; Epand, R M

2000-12-01

A regular progression of polymorphic phase behavior was observed for mixtures of the anionic phospholipid, cardiolipin, and the cationic phospholipid derivative, 1, 2-dioleoyl-sn-glycero-3-ethylphosphocholine. As revealed by freeze-fracture electron microscopy and small-angle x-ray diffraction, whereas the two lipids separately assume only lamellar phases, their mixtures exhibit a symmetrical (depending on charge ratio and not polarity) sequence of nonlamellar phases. The inverted hexagonal phase, H(II,) formed from equimolar mixtures of the two lipids, i.e., at net charge neutrality (charge ratio (CR((+/-))) = 1:1). When one type of lipid was in significant excess (CR((+/-)) = 2:1 or CR((+/-)) = 1:2), a bicontinuous cubic structure was observed. These cubic phases were very similar to those sometimes present in cellular organelles that contain cardiolipin. Increasing the excess of cationic or anionic charge to CR((+/-)) = 4:1 or CR((+/-)) = 1:4 led to the appearance of membrane bilayers with numerous interlamellar contacts, i.e., sponge structures. It is evident that interactions between cationic and anionic moieties can influence the packing of polar heads and hence control polymorphic phase transitions. The facile isothermal, polymorphic interconversion of these lipids may have important biological and technical implications.

Biomimetic mineralization of CaCO3 on a phospholipid monolayer: from an amorphous calcium carbonate precursor to calcite via vaterite.

Science.gov (United States)

Xiao, Junwu; Wang, Zhining; Tang, Yecang; Yang, Shihe

2010-04-06

A phospholipid monolayer, approximately half the bilayer structure of a biological membrane, can be regarded as an ideal model for investigating biomineralization on biological membranes. In this work on the biomimetic mineralization of CaCO(3) under a phospholipid monolayer, we show the initial heterogeneous nucleation of amorphous calcium carbonate precursor (ACC) nanoparticles at the air-water interface, their subsequent transformation into the metastable vaterite phase instead of the most thermodynamically stable calcite phase, and the ultimate phase transformation to calcite. Furthermore, the spontaneity of the transformation from vaterite to calcite was found to be closely related to the surface tension; high surface pressure could inhibit the process, highlighting the determinant of surface energy. To understand better the mechanisms for ACC formation and the transformation from ACC to vaterite and to calcite, in situ Brewster angle microscopy (BAM), ex situ scanning electron microscopy, transmission electron microscopy, Raman spectroscopy, and X-ray diffraction analysis were employed. This work has clarified the crystallization process of calcium carbonate under phospholipid monolayers and therefore may further our understanding of the biomineralization processes induced by cellular membranes.

Metabolism of fatty acids in rat brain in microsomal membranes

International Nuclear Information System (INIS)

Aeberhard, E.E.; Gan-Elepano, M.; Mead, J.F.

1980-01-01

Using a technique in which substrate fatty acids are incorporated into microsomal membranes followd by comparison of their rates of desaturation or elongation with those of exogenous added fatty acids it has been found that the desaturation rate is more rapid for the membrane-bound substrate than for the added fatty acid. Moreover, the product of the membrane-bound substrate is incorporated into membrane phospholipid whereas the product of the exogenous substrate is found in di- and triacyl glycerols and in free fatty acids as well. These and other findings point to a normal sequence of reaction of membrane liqids with membrane-bound substrates involving transfer of fatty acid from phospholipid to the coupled enzyme systems without ready equilibration with the free fatty acid pool

Using neurolipidomics to identify phospholipid mediators of synaptic (dysfunction in Alzheimer’s Disease

Directory of Open Access Journals (Sweden)

Steffany A L Bennett

2013-07-01

Full Text Available Not all of the mysteries of life lie in our genetic code. Some can be found buried in our membranes. These shells of fat, sculpted in the central nervous system into the cellular (and subcellular boundaries of neurons and glia, are themselves complex systems of information. The diversity of neural phospholipids, coupled with their chameleon-like capacity to transmute into bioactive molecules, provides a vast repertoire of immediate response second messengers. The effects of compositional changes on synaptic function have only begun to be appreciated. Here, we mined 29 different neurolipidomic datasets for changes in neuronal membrane phospholipid metabolism in Alzheimer’s Disease. Three overarching metabolic disturbances were detected. We found that an increase in the hydrolysis of platelet activating factor precursors and ethanolamine-containing plasmalogens, coupled with a failure to regenerate relatively rare alkyl-acyl and alkenyl-acyl structural phospholipids, correlated with disease severity. Accumulation of specific bioactive metabolites (i.e., PC(O-16:0/2:0 and PE(P-16:0/0:0 was associated with aggravating tau pathology, enhancing vesicular release, and signaling neuronal loss. Finally, depletion of PI(16:0/20:4, PI(16:0/22:6, and PI(18:0/22:6 was implicated in accelerating Aβ42 biogenesis. Our analysis further suggested that converging disruptions in platelet activating factor, plasmalogen, phosphoinositol and phosphoethanolamine, and docosahexaenoic acid metabolism may contribute mechanistically to catastrophic vesicular depletion, impaired receptor trafficking, and morphological dendritic deformation. Together, this analysis supports an emerging hypothesis that aberrant phospholipid metabolism may be one of multiple critical determinants required for Alzheimer disease conversion.

13C-labeled 18 : 2n-6 recovered in brush border membrane phospholipids short time after administration

DEFF Research Database (Denmark)

Vistisen, Bodil; Høy, Carl-Erik

2004-01-01

fatty acids in the two phospholipid pools. Minor effects on BBM-PC and BBM-PE fatty acid profiles (mole-%) were observed. The present study demonstrated for the first time incorporation of C-13-labeled 18:2n-6 into BBM-PC 2 hours and 6 hours after intragastric administration of L*L*L* or ML......*M. This emphasizes the influence of the dietary fatty acid on BBM fatty acid composition and the rapid incorporation of dietary long-chain fatty acids into intestinal enterocyte phospholipids. Medium-chain fatty acids in a single meal exert only a minor influence on the BBM phospholipid fatty acid profile....

Chemistry of phospholipid oxidation.

Science.gov (United States)

Reis, Ana; Spickett, Corinne M

2012-10-01

The oxidation of lipids has long been a topic of interest in biological and food sciences, and the fundamental principles of non-enzymatic free radical attack on phospholipids are well established, although questions about detail of the mechanisms remain. The number of end products that are formed following the initiation of phospholipid peroxidation is large, and is continually growing as new structures of oxidized phospholipids are elucidated. Common products are phospholipids with esterified isoprostane-like structures and chain-shortened products containing hydroxy, carbonyl or carboxylic acid groups; the carbonyl-containing compounds are reactive and readily form adducts with proteins and other biomolecules. Phospholipids can also be attacked by reactive nitrogen and chlorine species, further expanding the range of products to nitrated and chlorinated phospholipids. Key to understanding the mechanisms of oxidation is the development of advanced and sensitive technologies that enable structural elucidation. Tandem mass spectrometry has proved invaluable in this respect and is generally the method of choice for structural work. A number of studies have investigated whether individual oxidized phospholipid products occur in vivo, and mass spectrometry techniques have been instrumental in detecting a variety of oxidation products in biological samples such as atherosclerotic plaque material, brain tissue, intestinal tissue and plasma, although relatively few have achieved an absolute quantitative analysis. The levels of oxidized phospholipids in vivo is a critical question, as there is now substantial evidence that many of these compounds are bioactive and could contribute to pathology. The challenges for the future will be to adopt lipidomic approaches to map the profile of oxidized phospholipid formation in different biological conditions, and relate this to their effects in vivo. This article is part of a Special Issue entitled: Oxidized phospholipids

Transport of anhydrous ammoniac - risk assessment; Le transport de l'ammoniac anhydre - analyse et estimation des risques: rapport de synthese

Energy Technology Data Exchange (ETDEWEB)

Castellano, S; Degrange, J P; Hubert, P; Pages, P; Lamblin, J

1989-12-01

This risk analysis of anhydrous ammonia transport in France was done within a study initiated by the Department of dangerous goods of the Ministry of transport. The study deals with the road and rail transportation of bulk anhydrous ammonia. After analysis of transport system and traffic, the transport accident risks are estimated, as well as their distribution on the French territory. Finally after a synthesis of results, a number of safety measures to be undertaken were identified. This is a joint study of SMC-CEPN, with a specific role of SEMA-METRA-CONSEIL concerning the traffic frequency, and the center for risk evaluation concerning nuclear safety.

Membrane phospholipids and radiation-induced death of mammalian cells

International Nuclear Information System (INIS)

Wolters, H.

1987-01-01

Radiation-induced cell killing is generally believed to be a consequence of residual DNA damage or damage that is mis-repaired. However, besides this DNA damage, damage to other molecules or structures of the cell may be involved in the killing. Especially membranes have been suggested as a determinant in cellular radiosensitivity. In this thesis experiments are described, dealing with the possible involvement of membranes in radiation-induced killing of mammalian cells. A general treatise of membrane structure is followed by information concerning deleterious effects of radiation on membranes. Consequences of damage to structure and function of membranes are reviewed. Thereafter evidence relating to the possible involvement of membranes in radiation-induced cell killing is presented. (Auth.)

Phospholipids and protein adaptation of Pseudomonas sp. to the xenoestrogen tributyltin chloride (TBT).

Science.gov (United States)

Bernat, Przemysław; Siewiera, Paulina; Soboń, Adrian; Długoński, Jerzy

2014-09-01

A tributyltin (TBT)-resistant strain of Pseudomonas sp. isolated from an overworked car filter was tested for its adaptation to TBT. The isolate was checked for organotin degradation ability, as well as membrane lipid and cellular protein composition in the presence of TBT. The phospholipid profiles of bacteria, grown with and without increased amounts of TBT, were characterized using liquid chromatography/electrospray ionization/mass spectrometry. The strain reacted to the biocide by changing the composition of its phospholipids. TBT induced a twofold decline in the amounts of many molecular species of phosphatidylglycerol and an increase in the levels of phosphatidic acid (by 58%) and phosphatidylethanolamine (by 70%). An increase in the degree of saturation of phospholipid fatty acids of TBT exposed Pseudomonas sp. was observed. These changes in the phospholipid composition and concentration reflect the mechanisms which support optimal lipid ordering in the presence of toxic xenobiotic. In the presence of TBT the abundances of 16 proteins, including TonB-dependent receptors, porins and peroxidases were modified, which could indicate a contribution of some enzymes to TBT resistance.

Increased Binding of Calcium Ions at Positively Curved Phospholipid Membranes

Czech Academy of Sciences Publication Activity Database

Magarkar, Aniket; Jurkiewicz, Piotr; Allolio, Christoph; Hof, Martin; Jungwirth, Pavel

2017-01-01

Roč. 8, č. 2 (2017), s. 518-523 ISSN 1948-7185 R&D Projects: GA ČR(CZ) GA16-01074S; GA ČR(CZ) GAP207/12/0919 Grant - others:AV ČR(CZ) AP1102 Program:Akademická prémie - Praemium Academiae Institutional support: RVO:61388963 ; RVO:61388955 Keywords : molecular dynamics * fluorescence spectroscopy * calcium * phospholipids Subject RIV: CF - Physical ; Theoretical Chemistry; CF - Physical ; Theoretical Chemistry (UFCH-W) OBOR OECD: Physical chemistry; Physical chemistry (UFCH-W) Impact factor: 9.353, year: 2016

Phospholipid Binding Protein C Inhibitor (PCI) Is Present on Microparticles Generated In Vitro and In Vivo

Science.gov (United States)

Einfinger, Katrin; Badrnya, Sigrun; Furtmüller, Margareta; Handschuh, Daniela; Lindner, Herbert; Geiger, Margarethe

2015-01-01

Protein C inhibitor is a secreted, non-specific serine protease inhibitor with broad protease reactivity. It binds glycosaminoglycans and anionic phospholipids, which can modulate its activity. Anionic phospholipids, such as phosphatidylserine are normally localized to the inner leaflet of the plasma membrane, but are exposed on activated and apoptotic cells and on plasma membrane-derived microparticles. In this report we show by flow cytometry that microparticles derived from cultured cells and activated platelets incorporated protein C inhibitor during membrane blebbing. Moreover, protein C inhibitor is present in/on microparticles circulating in normal human plasma as judged from Western blots, ELISAs, flow cytometry, and mass spectrometry. These plasma microparticles are mainly derived from megakaryocytes. They seem to be saturated with protein C inhibitor, since they do not bind added fluorescence-labeled protein C inhibitor. Heparin partially removed microparticle-bound protein C inhibitor, supporting our assumption that protein C inhibitor is bound via phospholipids. To assess the biological role of microparticle-bound protein C inhibitor we performed protease inhibition assays and co-precipitated putative binding partners on microparticles with anti-protein C inhibitor IgG. As judged from amidolytic assays microparticle-bound protein C inhibitor did not inhibit activated protein C or thrombin, nor did microparticles modulate the activity of exogenous protein C inhibitor. Among the proteins co-precipitating with protein C inhibitor, complement factors, especially complement factor 3, were most striking. Taken together, our data do not support a major role of microparticle-associated protein C inhibitor in coagulation, but rather suggest an interaction with proteins of the complement system present on these phospholipid vesicles. PMID:26580551

ER phospholipid composition modulates lipogenesis during feeding and in obesity.

Science.gov (United States)

Rong, Xin; Wang, Bo; Palladino, Elisa Nd; de Aguiar Vallim, Thomas Q; Ford, David A; Tontonoz, Peter

2017-10-02

Sterol regulatory element-binding protein 1c (SREBP-1c) is a central regulator of lipogenesis whose activity is controlled by proteolytic cleavage. The metabolic factors that affect its processing are incompletely understood. Here, we show that dynamic changes in the acyl chain composition of ER phospholipids affect SREBP-1c maturation in physiology and disease. The abundance of polyunsaturated phosphatidylcholine in liver ER is selectively increased in response to feeding and in the setting of obesity-linked insulin resistance. Exogenous delivery of polyunsaturated phosphatidylcholine to ER accelerated SREBP-1c processing through a mechanism that required an intact SREBP cleavage-activating protein (SCAP) pathway. Furthermore, induction of the phospholipid-remodeling enzyme LPCAT3 in response to liver X receptor (LXR) activation promoted SREBP-1c processing by driving the incorporation of polyunsaturated fatty acids into ER. Conversely, LPCAT3 deficiency increased membrane saturation, reduced nuclear SREBP-1c abundance, and blunted the lipogenic response to feeding, LXR agonist treatment, or obesity-linked insulin resistance. Desaturation of the ER membrane may serve as an auxiliary signal of the fed state that promotes lipid synthesis in response to nutrient availability.

1,2-Dielaidoylphosphocholine/1,2-dimyristoylphosphoglycerol supported phospholipid bilayer formation in calcium and calcium-free buffer

International Nuclear Information System (INIS)

Evans, Kervin O.

2012-01-01

Phospholipid membranes are useful in the field of biocatalysis because a supported phospholipid membrane can create a biomimetic platform where biocatalytic processes can readily occur. In this work, supported bilayer formation from sonicated phospholipid vesicles containing 1,2-dielaidoyl-sn-glycero-3-phosphocholine and 1,2-dimyristoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] was studied using a quartz crystal microbalance with dissipation monitoring and an atomic force microscope. The molar percentages of DEPC and DMPG were varied to determine the effect of overall lipid composition on supported bilayer formation. This work also explored the effect that calcium ion concentration had on supported bilayer formation. Results show that vesicles with up to 50 mol% dimyristoylphosphoglycerol can form a supported bilayer without the presence of calcium ions; however, supported bilayer formation in calcium buffer was inhibited as the anionic (negatively charged) lipid concentration increased. Data suggest that supported phospholipid bilayer formation in the absence of Ca 2+ from vesicles containing negatively charged lipids is specific to phosphatidylglycerol. - Highlights: ► SPB formation of DEPC vesicles containing 0 to 50 mol% DMPG monitored using QCM-D. ► Ca 2+ inhibited SPB formation of DEPC vesicles containing 30 to 50 mol% DMPG. ► Vesicles containing DMPG at 0 to 50 mol% formed SPB in buffer free of Ca 2+ .

A novel isolation strategy for obtaining crude membrane vesicles from bovine skim milk

DEFF Research Database (Denmark)

Blans, Kristine; Larsen, Lotte Bach; Wiking, Lars

Bovine milks content of phospholipid membranes have largely been explored in the cream fraction, and known as the milk fat globule membrane that surrounds fat droplets. In skim milk, the population of phospholipid membranes is reported to constitute membrane vesicles with a soluble content known...... is observed all over the gradient. The variety of the membrane vesicles is currently being investigated further by several means. Summary/conclusion: A new procedure for easy and gentle isolation of bovine milk membrane vesicles encompassing ultracentrifugation and size-exclusion chromatography has been...... established. The resulting vesicle isolate exhibits the general membrane vesicle characteristics and provides an appropriate start material from which the variety of milk vesicles can be investigated...

Membrane phosphorylation and nerve cell function

International Nuclear Information System (INIS)

Baer, P.R.

1982-01-01

This thesis deals with the phosphorylation of membrane components. In part I a series of experiments is described using the hippocampal slice as a model system. In part II a different model system - cultured hybrid cells - is used to study protein and lipid phosphorylation, influenced by incubation with neuropeptides. In part III in vivo and in vitro studies are combined to study protein phosphorylation after neuroanatomical lesions. In a section of part II (Page 81-90) labelling experiments of the membrane inositol-phospholipids are described. 32 P-ATP was used to label phospholipids in intact hybrid cells, and short incubations were found to be the most favourable. (C.F.)

Field-effect detection using phospholipid membranes -Topical Review

Directory of Open Access Journals (Sweden)

Chiho Kataoka-Hamai and Yuji Miyahara

2010-01-01

Full Text Available The application of field-effect devices to biosensors has become an area of intense research interest. An attractive feature of field-effect sensing is that the binding or reaction of biomolecules can be directly detected from a change in electrical signals. The integration of such field-effect devices into cell membrane mimics may lead to the development of biosensors useful in clinical and biotechnological applications. This review summarizes recent studies on the fabrication and characterization of field-effect devices incorporating model membranes. The incorporation of black lipid membranes and supported lipid monolayers and bilayers into semiconductor devices is described.

Apolipoprotein L1 confers pH-switchable ion permeability to phospholipid vesicles.

Science.gov (United States)

Bruno, Jonathan; Pozzi, Nicola; Oliva, Jonathan; Edwards, John C

2017-11-03

Apolipoprotein L1 (ApoL1) is a human serum protein conferring resistance to African trypanosomes, and certain ApoL1 variants increase susceptibility to some progressive kidney diseases. ApoL1 has been hypothesized to function like a pore-forming colicin and has been reported to have permeability effects on both intracellular and plasma membranes. Here, to gain insight into how ApoL1 may function in vivo , we used vesicle-based ion permeability, direct membrane association, and intrinsic fluorescence to study the activities of purified recombinant ApoL1. We found that ApoL1 confers chloride-selective permeability to preformed phospholipid vesicles and that this selectivity is strongly pH-sensitive, with maximal activity at pH 5 and little activity above pH 7. When ApoL1 and lipid were allowed to interact at low pH and were then brought to neutral pH, chloride permeability was suppressed, and potassium permeability was activated. Both chloride and potassium permeability linearly correlated with the mass of ApoL1 in the reaction mixture, and both exhibited lipid selectivity, requiring the presence of negatively charged lipids for activity. Potassium, but not chloride, permease activity required the presence of calcium ions in both the association and activation steps. Direct assessment of ApoL1-lipid associations confirmed that ApoL1 stably associates with phospholipid vesicles, requiring low pH and the presence of negatively charged phospholipids for maximal binding. Intrinsic fluorescence of ApoL1 supported the presence of a significant structural transition when ApoL1 is mixed with lipids at low pH. This pH-switchable ion-selective permeability may explain the different effects of ApoL1 reported in intracellular and plasma membrane environments. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Lipids in the Assembly of Membrane Proteins and Organization of Protein Supercomplexes: Implications for Lipid-Linked Disorders

OpenAIRE

Bogdanov, Mikhail; Mileykovskaya, Eugenia; Dowhan, William

2008-01-01

Lipids play important roles in cellular dysfunction leading to disease. Although a major role for phospholipids is in defining the membrane permeability barrier, phospholipids play a central role in a diverse range of cellular processes and therefore are important factors in cellular dysfunction and disease. This review is focused on the role of phospholipids in normal assembly and organization of the membrane proteins, multimeric protein complexes, and higher order supercomplexes. Since lipi...

Phosphoenolpyruvate carboxylase from C4 leaves is selectively targeted for inhibition by anionic phospholipids

NARCIS (Netherlands)

Monreal, J.A.; McLoughlin, F.; Echevarría, C.; García-Mauriño, S.; Testerink, C.

2010-01-01

Phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31) is an enzyme playing a crucial role in photosynthesis of C4 plants. Here, we identify anionic phospholipids as novel regulators that inhibit C4 PEPC activity and provide evidence that the enzyme partially localizes to membranes.

Study of alkaline-earth element complexes in anhydrous acetic acid

International Nuclear Information System (INIS)

Petit, N.

1968-10-01

We have studied the complexes of alkaline-earth elements in anhydrous acetic acid. Using glass-electrode potentiometry we have studied the titration of alkaline earth acetates with perchloric acid which is the strongest acid in anhydrous acetic acid. These titrations have shown that the basic strength of these acetates increases as follows: Mg 4 ); the mixed acetate-acid sulfate complex of barium: Ba (OAc)(HSO 4 ); the mixed acetate-chloride of barium: Ba (OAc)(Cl). (author) [fr

Training affects muscle phospholipid fatty acid composition in humans

DEFF Research Database (Denmark)

Helge, Jørn Wulff; Wu, B J; Willer, Mette

2001-01-01

on the muscle membrane phospholipid fatty acid composition in humans. Seven male subjects performed endurance training of the knee extensors of one leg for 4 wk. The other leg served as a control. Before, after 4 days, and after 4 wk, muscle biopsies were obtained from the vastus lateralis. After 4 wk......, the phospholipid fatty acid contents of oleic acid 18:1(n-9) and docosahexaenoic acid 22:6(n-3) were significantly higher in the trained (10.9 +/- 0.5% and 3.2 +/- 0.4% of total fatty acids, respectively) than the untrained leg (8.8 +/- 0.5% and 2.6 +/- 0.4%, P fatty acids...... was significantly lower in the trained (11.1 +/- 0.9) than the untrained leg (13.1 +/- 1.2, P fatty acid composition. Citrate synthase activity was increased by 17% in the trained compared with the untrained leg (P

Perforin rapidly induces plasma membrane phospholipid flip-flop.

Directory of Open Access Journals (Sweden)

Sunil S Metkar

Full Text Available The cytotoxic cell granule secretory pathway is essential for host defense. This pathway is fundamentally a form of intracellular protein delivery where granule proteases (granzymes from cytotoxic lymphocytes are thought to diffuse through barrel stave pores generated in the plasma membrane of the target cell by the pore forming protein perforin (PFN and mediate apoptotic as well as additional biological effects. While recent electron microscopy and structural analyses indicate that recombinant PFN oligomerizes to form pores containing 20 monomers (20 nm when applied to liposomal membranes, these pores are not observed by propidium iodide uptake in target cells. Instead, concentrations of human PFN that encourage granzyme-mediated apoptosis are associated with pore structures that unexpectedly favor phosphatidylserine flip-flop measured by Annexin-V and Lactadherin. Efforts that reduce PFN mediated Ca influx in targets did not reduce Annexin-V reactivity. Antigen specific mouse CD8 cells initiate a similar rapid flip-flop in target cells. A lipid that augments plasma membrane curvature as well as cholesterol depletion in target cells enhance flip-flop. Annexin-V staining highly correlated with apoptosis after Granzyme B (GzmB treatment. We propose the structures that PFN oligomers form in the membrane bilayer may include arcs previously observed by electron microscopy and that these unusual structures represent an incomplete mixture of plasma membrane lipid and PFN oligomers that may act as a flexible gateway for GzmB to translocate across the bilayer to the cytosolic leaflet of target cells.

Encapsulation of phytosterols and phytosterol esters in liposomes made with soy phospholipids by high pressure homogenization.

Science.gov (United States)

Wang, Fan C; Acevedo, Nuria; Marangoni, Alejandro G

2017-11-15

Phytosterols and phytosterol esters were encapsulated within large unilamellar liposomes prepared with soy phospholipids using a microfluidizer. The average particle diameter of these liposomal vesicles increased with increasing amounts of encapsulated phytosterols, especially with increasing free sterol content. The phytosterol content, liposomal particle size, and phytosterol encapsulation efficiency started to plateau when liposomes were prepared with MOPS buffer dispersions that contained 50 mg ml -1 soy phospholipid and more than 4% phytosterol blend, suggesting the saturation of phytosterol encapsulation. We proposed an encapsulation mechanism of free sterols and phytosterol esters in liposomes, where free sterols were mainly encapsulated within the lumen of these liposomes as crystals, and sterol esters and some free sterols were incorporated within the phospholipid bilayer of the liposomal membrane. The results from this work could provide the pharmaceutical and nutraceutical industries a practical method to produce loaded liposomes using inexpensive phospholipid mixtures for the delivery of bioactive ingredients.

Determination of the separate lipid and protein profile structures derived from the total membrane profile structure or isolated sarcoplasmic reticulum via x-ray and neutron diffraction

International Nuclear Information System (INIS)

Herbette, L.; Blasie, J.K.

1984-01-01

Sarcoplasmic reticulum (SR) membranes were prepared to contain biosynthetically deuterated SR phospholipids utilizing specific and general phospholipid exchange proteins (PLEP). Functional measurements and freeze fracture on SR dispersions and x-ray diffraction of hydrated oriented membrane multilayers revealed that the exchanged SR membranes were very similar to unexchanged SR membranes. Low resolution (28-A) neutron diffraction studies utilizing SR membranes exchanged with either protonated or perdeuterated SR phospholipids allowed direct determination of the lipid profile within the isolated SR membrane at two different unit cell repeat distances. These lipid profile structures were found to be highly asymmetric regarding the conformation of the fatty acid chain extents and compositional distribution of phospholipid molecules in the inner vs. outer monolayer of the SR membrane bilayer. The relatively high resolution (11-A) electron-density profile from x-ray diffraction was decomposed by utilizing the asymmetry in the number of phospholipid molecules residing in the inner vs. outer monolayer of the SR lipid bilayer as obtained from the neutron diffraction study. To our knowledge, this represents the first direct determination of a lipid bilayer profile structure within an isolated membrane system

Conformations of double-headed, triple-tailed phospholipid oxidation lipid products in model membranes

DEFF Research Database (Denmark)

Hermetter, Albin; Kopec, Wojciech; Khandelia, Himanshu

2013-01-01

Products of phospholipid oxidation can produce lipids with a carbonyl moiety at the end of a shortened lipid acyl tail, such as 1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphocholine (POVPC). The carbonyl tail of POVPC can covalently bond to the free tertiary amine of a phosphatidylethanolamine...

Formation of a new crystalline form of anhydrous β-maltose by ethanol-mediated crystal transformation.

Science.gov (United States)

Verhoeven, Nicolas; Neoh, Tze Loon; Ohashi, Tetsuya; Furuta, Takeshi; Kurozumi, Sayaka; Yoshii, Hidefumi

2012-04-01

β-Maltose monohydrate was transformed into an anhydrous form by ethanol-mediated method under several temperatures with agitation. A new stable anhydrous form of β-maltose (Mβ(s)) was obtained, as substantiated by the X-ray diffraction patterns. Mβ(s) obtained by this method presented a fine porous structure, resulting in greater specific surface area compared to those of β-maltose monohydrate and anhydrous β-maltose obtained by vacuum drying (Mβ(h)). The crystal transformation presumably consisted of two steps: dehydration reaction from the hydrous to amorphous forms and crystal formation from the amorphous forms to the noble anhydrous form. The kinetics of these reactions were determined by thermal analysis using Jander's equation and Arrhenius plots. The overall activation energies of the dehydration reaction and the formation of anhydrous maltose were evaluated to be 100 and 90 kJ/mol, respectively. Copyright © 2012 Elsevier Ltd. All rights reserved.

Structural basis for the transformation pathways of the sodium naproxen anhydrate-hydrate system

DEFF Research Database (Denmark)

Bond, Andrew; Cornett, Claus; Larsen, Flemming Hofmann

2014-01-01

to the monohydrate (MH) and anhydrate (AH) structures, provide a basis to rationalize the observed transformation pathways in the sodium (S)-naproxen anhydrate-hydrate system. All structures contain Na(+)/carboxylate/H2O sections, alternating with sections containing the naproxen molecules. The structure of DH...

Phospholipid layer stabilization via Yb(III) on ITIES and facilitated K(I) transport

Czech Academy of Sciences Publication Activity Database

Jaklová Dytrtová, Jana; Navrátil, Tomáš; Mareček, Vladimír

2011-01-01

Roč. 76, č. 12 (2011), s. 1917-1930 ISSN 0010-0765 R&D Projects: GA AV ČR IAA400400806; GA ČR GAP206/11/1638 Institutional research plan: CEZ:AV0Z40400503; CEZ:AV0Z40550506 Keywords : phospholipids * membranes * zwitterions Subject RIV: CG - Electrochemistry Impact factor: 1.283, year: 2011

THz characterization of hydrated and anhydrous materials

Science.gov (United States)

Sokolnikov, Andre

2011-06-01

The characterization of anhydrous and hydrated forms of materials is of great importance to science and industry. Water content poses difficulties for successful identification of the material structure by THz radiation. However, biological tissues and hydrated forms of nonorganic substances still may be investigated by THz radiation. This paper outlines the range of possibilities of the above characterization, as well as provides analysis of the physical mechanism that allows or prevents penetration of THz waves through the substance. THz-TDS is used to measure the parameters of the characterization of anhydrous and hydrated forms of organic and nonorganic samples. Mathematical methods (such as prediction models of time-series analysis) are used to help identifying the absorption coefficient and other parameters of interest. The discovered dependencies allow designing techniques for material identification/characterization (e.g. of drugs, explosives, etc. that may have water content). The results are provided.

Saturated fatty acid in the phospholipid monolayer contributes to the formation of large lipid droplets

International Nuclear Information System (INIS)

Arisawa, Kotoko; Mitsudome, Haruka; Yoshida, Konomi; Sugimoto, Shizuka; Ishikawa, Tomoko; Fujiwara, Yoko; Ichi, Ikuyo

2016-01-01

The degree of saturation of fatty acid chains in the bilayer membrane structure is known to control membrane fluidity and packing density. However, the significance of fatty acid composition in the monolayers of lipid droplets (LDs) has not been elucidated. In this study, we noted a relationship between the size of LDs and the fatty acid composition of the monolayer. To obtain large LDs, we generated NIH3T3 cells overexpressing fat-specific protein 27 (FSP27). This induced the fusion of LDs, resulting in larger LDs in FSP27-overexpressing cells compared with LDs in control cells. Moreover, the lipid extracts of LDs from FSP27-overexpressing cells reconstituted large-droplet emulsions in vitro, implying that the lipid properties of LDs might affect the size of LDs. FSP27-overexpressing cells had more saturated fatty acids in the phospholipid monolayer of the LDs compared with control cells. To further investigate the effects of the degree of phospholipid unsaturation on the size of LDs, we synthesized artificial emulsions of a lipid mixed with distearoylphosphatidylcholine (DSPC, diC18:0-PC) and with dioleoylphosphatidylcholine (DOPC, diC18:1n-9-PC) and compared the sizes of the resulting LDs. The emulsions prepared from saturated PC had larger droplets than those prepared from unsaturated PC. Our results suggest that saturated fatty acid chains in phospholipid monolayers might establish the form and/or stability of large LDs. - Highlights: • The lipid extracts of larger LDs from FSP27 cells reconstructed large-droplet emulsions. • Isolated LDs from FSP27 cells had more saturated fatty acids in the phospholipid monolayer compared with the control. • Saturated fatty acids in the phospholipid monolayer are a factor in the formation of large emulsions.

pH-Induced Changes in the Surface Viscosity of Unsaturated Phospholipids Monitored Using Active Interfacial Microrheology.

Science.gov (United States)

Ghazvini, Saba; Alonso, Ryan; Alhakamy, Nabil; Dhar, Prajnaparamita

2018-01-23

Lipid membranes, a major component of cells, are subjected to significant changes in pH depending on their location in the cell: the outer leaflet of the cell membrane is exposed to a pH of 7.4 whereas lipid membranes that make up late endosomes and lysosomes are exposed to a pH of as low as 4.4. The purpose of this study is to evaluate how changes in the environmental pH within cells alter the fluidity of phospholipid membranes. Specifically, we studied pH-induced alterations in the surface arrangement of monounsaturated lipids with zwitterionic headgroups (phosphoethanolamine (PE) and phosphocholine (PC)) that are abundant in plasma membranes as well as anionic lipids (phosphatidylserine (PS) and phosphatidylglycerol (PG)) that are abundant in inner membranes using a combination of techniques including surface tension vs area measurements, interfacial microrheology, and fluorescence/atomic force microscopy. Using an active interfacial microrheology technique, we find that phospholipids with zwitterionic headgroups show a significant increase in their surface viscosity at acidic pH. This increase in surface viscosity is also found to depend on the size of the lipid headgroup, with a smaller headgroup showing a greater increase in viscosity. The observed pH-induced increase in viscosity is also accompanied by an increase in the cohesion pressure between zwitterionic molecules at acidic pH and a decrease in the average molecular area of the lipids, as measured by fitting the surface pressure isotherms to well-established equations of state. Because fluorescent images show no change in the phase of the lipids, we attribute this change in surface viscosity to the pH-induced reorientation of the P - -N + dipoles that form part of the polar lipid headgroup, resulting in increased lipid-lipid interactions. Anionic PG headgroups do not demonstrate this pH-induced change in viscosity, suggesting that the presence of a net negative charge on the headgroup causes

Forms, Crosstalks, and the Role of Phospholipid Biosynthesis in Autophagy

Directory of Open Access Journals (Sweden)

Leanne Pereira

2012-01-01

Full Text Available Autophagy is a highly conserved cellular process occurring during periods of stress to ensure a cell's survival by recycling cytosolic constituents and making products that can be used in energy generation and other essential processes. Three major forms of autophagy exist according to the specific mechanism through which cytoplasmic material is transported to a lysosome. Chaperone-mediated autophagy is a highly selective form of autophagy that delivers specific proteins for lysosomal degradation. Microautophagy is a less selective form of autophagy that occurs through lysosomal membrane invaginations, forming tubes and directly engulfing cytoplasm. Finally, macroautophagy involves formation of new membrane bilayers (autophagosomes that engulf cytosolic material and deliver it to lysosomes. This review provides new insights on the crosstalks between different forms of autophagy and the significance of bilayer-forming phospholipid synthesis in autophagosomal membrane formation.

Anhydrous hydrogen fluoride electrolyte battery. [Patent application

Science.gov (United States)

Not Available

1972-06-26

It is an object of the invention to provide a primary cell or battery using ammonium fluoride--anhydrous hydrogen fluoride electrolyte having improved current and power production capabilities at low temperatures. It is operable at temperatures substantially above the boiling point of hydrogen fluoride. (GRA)

Enhancement of proton transfer in ion channels by membrane phosphate headgroups.

Science.gov (United States)

Wyatt, Debra L; de Godoy, Carlos Marcelo G; Cukierman, Samuel

2009-05-14

The transfer of protons (H+) in gramicidin (gA) channels is markedly distinct in monoglyceride and phospholipid membranes. In this study, the molecular groups that account for those differences were investigated using a new methodology. The rates of H+ transfer were measured in single gA channels reconstituted in membranes made of plain ceramides or sphingomyelins and compared to those in monoglyceride and phospholipid bilayers. Single-channel conductances to protons (gH) were significantly larger in sphingomyelin than in ceramide membranes. A novel and unsuspected finding was that H+ transfer was heavily attenuated or completely blocked in ceramide (but not in sphingomyelin) membranes in low-ionic-strength solutions. It is reasoned that H-bond dynamics at low ionic strengths between membrane ceramides and gA makes channels dysfunctional. The rate of H+ transfer in gA channels in ceramide membranes is significantly higher than that in monoglyceride bilayers. This suggests that solvation of the hydrophobic surface of gA channels by two acyl chains in ceramides stabilizes the gA channels and the water wire inside the pore, leading to an enhancement of H+ transfer in relation to that occurring in monoglyceride membranes. gH values in gA channels are similar in ceramide and monoglyceride bilayers and in sphingomyelin and phospholipid membranes. It is concluded that phospho headgroups in membranes have significant effects on the rate of H+ transfer at the membrane gA channel/solution interfaces, enhancing the entry and exit rates of protons in channels.

Insulin and adenosine regulate the phosphatidylcholine concentration in isolated rat adipocyte plasma membranes.

Science.gov (United States)

Kiechle, F L; Sykes, E; Artiss, J D

1995-01-01

Blockade of adenosine receptors by 3-isobutyl-1-methylxanthine or degradation of endogenous adenosine with adenosine deaminase increased the phosphatidylcholine concentration in isolated rat adipocyte plasma membranes, an effect which was suppressed by the phosphatidylethanolamine methyltransferase inhibitor, S-adenosyl-L-homocysteine, and reversed by the adenosine analogue, N6-(L-phenylisopropyl)-adenosine. For example, the addition of N6-(L-phenylisopropyl)-adenosine to adenosine deaminase pretreated plasma membranes rapidly lowered the concentration of phosphatidylcholine by 171 nmol/mg at 30 seconds compared to control. Insulin-induced stimulation of phospholipid methylation in membranes treated with 3-isobutyl-1-methylxanthine or adenosine deaminase was achieved only after the addition of N6-(L-phenylisopropyl)-adenosine. These results suggest that adenosine receptor occupancy inhibits phospholipid methylation, is required for insulin stimulation of phospholipid methylation, and may perhaps activate a phosphatidylcholine-specific phospholipase C or phospholipase D.

Intermolecular crosslinks mediate aggregation of phospholipid vesicles by pulmonary surfactant-associated protein SAP-35

International Nuclear Information System (INIS)

Ross, G.R.; Sawyer, J.; Whitsett, J.

1987-01-01

Pulmonary surfactant-associated protein, Mr=35,000 (SAP-35) is known to bind phospholipids and is hypothesized to function in the organization of surfactant lipid membranes. SAP-35 has been observed to accelerate the calcium-induced aggregation of phospholipid vesicles. In order to define the molecular domains of SAP-35 which function in phospholipid aggregation, they have measured the light scattering properties (400nm) of purified canine SAP-35-phospholipid vesicle suspensions. Accelerated aggregation of unilamellar vesicles, requires SAP-35 and at least 2mM free calcium. The initial rate of A 400 change is proportional to the amount of native SAP-35 added over lipid:protein molar ratios ranging from 100:1 to 5000:1. Removal of the SAP-35 collagen-like domain and a specific cysteine residue involved in intermolecular disulfide bonding by bacterial collagenase digestion destroys the protein's lipid aggregation activity. Pre-incubation of SAP-35 with dithiothreitol (DTT) under nondenaturing conditions also results in a time-dependent loss of aggregation activity. Sucrose density gradient floatation of SAP-35 with 14 C dipalmitoyl phosphatidycholine labelled vesicles in the absence or presence of DTT suggests retention of SAP-35 lipid binding capacity. These data demonstrate the importance of SAP-35 triple helix and disulfide crosslinking integrity for the aggregation of unilamellar phospholipid vesicles

Annexin A4 and A6 induce membrane curvature and constriction during cell membrane repair

DEFF Research Database (Denmark)

Boye, Theresa Louise; Maeda, Kenji; Pezeshkian, Weria

2017-01-01

Efficient cell membrane repair mechanisms are essential for maintaining membrane integrity and thus for cell life. Here we show that the Ca2+- and phospholipid-binding proteins annexin A4 and A6 are involved in plasma membrane repair and needed for rapid closure of micron-size holes. We demonstrate...... that annexin A4 binds to artificial membranes and generates curvature force initiated from free edges, whereas annexin A6 induces constriction force. In cells, plasma membrane injury and Ca2+ influx recruit annexin A4 to the vicinity of membrane wound edges where its homo-trimerization leads to membrane...... that induction of curvature force around wound edges is an early key event in cell membrane repair....

Phospholipid composition of Dipylidium caninum.

Science.gov (United States)

Chopra, A K; Jain, S K; Vinayak, V K; Khuller, G K

1978-11-15

The phospholipid composition of Dipylidium caninum has been studied. Chloroform-methanol-soluble fraction amounted to 2.4% and phospholipids to 0.5% of the wet weight of the parasite. Phosphatidyl choline and phosphatidyl ethanolamine represented the bulk of the phospholipids, whereas phosphatidyl serine, phosphatidyl inositol, lysolecithin and lysophosphatidyl ethanolamine were present in minor amounts. Sulfatides were also identified in this parasite.

Progress in surface and membrane science

CERN Document Server

Cadenhead, D A

1976-01-01

Progress in Surface and Membrane Science, Volume 10 covers the advances in surface and membrane science. The book discusses the selective changes of cellular particles influencing sedimentation properties; and the rotating disk and ring-disk electrodes in investigations of surface phenomena at the metal-electrolyte interface. The text also describes the membrane potential of phospholipid bilayer and biological membranes; the adsorption of surfactant monolayers at gas/liquid and liquid/liquid interfaces; and the enzymes immobilized on glass. Chemists and people involved in electrochemistry will

Progress in surface and membrane science

CERN Document Server

Danielli, J F; Cadenhead, D A

1972-01-01

Progress in Surface and Membrane Science, Volume 5 covers the developments in the study of surface and membrane science. The book discusses the Mössbauer effect in surface science; the surface functional groups on carbon and silica; and the wetting phenomena pertaining to adhesion. The text also describes the physical state of phospholipids and cholesterol in monolayers, bilayers, and membranes; the characteristics of heterocoagulation; and the effects of calcium on excitable membranes and neurotransmitter action. Chemists, physiologists, biophysicists, and civil engineers will find the book i

The Physical Properties of Ceramides in Membranes.

Science.gov (United States)

Alonso, Alicia; Goñi, Félix M

2018-05-20

Ceramides are sphingolipids containing a sphingosine or a related base, to which a fatty acid is linked through an amide bond. When incorporated into a lipid bilayer, ceramides exhibit a number of properties not shared by almost any other membrane lipid: Ceramides ( a) are extremely hydrophobic and thus cannot exist in suspension in aqueous media; ( b) increase the molecular order (rigidity) of phospholipids in membranes; ( c) give rise to lateral phase separation and domain formation in phospholipid bilayers; ( d) possess a marked intrinsic negative curvature that facilitates formation of inverted hexagonal phases; ( e) make bilayers and cell membranes permeable to small and large (i.e., protein-size) solutes; and ( f) promote transmembrane (flip-flop) lipid motion. Unfortunately, there is hardly any link between the physical studies reviewed here and the mass of biological and clinical studies on the effects of ceramides in health and disease.

Polycyclic aromatic hydrocarbons in model bacterial membranes - Langmuir monolayer studies.

Science.gov (United States)

Broniatowski, Marcin; Binczycka, Martyna; Wójcik, Aneta; Flasiński, Michał; Wydro, Paweł

2017-12-01

High molecular weight polycyclic aromatic hydrocarbons (HMW-PAHs) are persistent organic pollutants which due to their limited biodegradability accumulate in soils where their increased presence can lead to the impoverishment of the decomposer organisms. As very hydrophobic PAHs easily penetrate cellular membranes of soil bacteria and can be incorporated therein, changing the membrane fluidity and other functions which in consequence can lead to the death of the organism. The structure and size of PAH molecule can be crucial for its membrane activity; however the correlation between PAH structure and its interaction with phospholipids have not been investigated so far. In our studies we applied phospholipid Langmuir monolayers as model bacterial membranes and investigated how the incorporation of six structurally different PAH molecules change the membrane texture and physical properties. In our studies we registered surface pressure and surface potential isotherms upon the monolayer compression, visualized the monolayer texture with the application of Brewster angle microscopy and searched the ordering of the film-forming molecules with molecular resolution with the application of grazing incidence X-ray diffraction (GIXD) method. It turned out that the phospholipid-PAH interactions are strictly structure dependent. Four and five-ring PAHs of the angular or cluster geometry can be incorporated into the model membranes changing profoundly their textures and fluidity; whereas linear or large cluster PAHs cannot be incorporated and separate from the lipid matrix. The observed phenomena were explained based on structural similarities of the applied PAHs with membrane steroids and hopanoids. Copyright © 2017. Published by Elsevier B.V.

A mitochondrially targeted compound delays aging in yeast through a mechanism linking mitochondrial membrane lipid metabolism to mitochondrial redox biology

Directory of Open Access Journals (Sweden)

Michelle T. Burstein

2014-01-01

Full Text Available A recent study revealed a mechanism of delaying aging in yeast by a natural compound which specifically impacts mitochondrial redox processes. In this mechanism, exogenously added lithocholic bile acid enters yeast cells, accumulates mainly in the inner mitochondrial membrane, and elicits an age-related remodeling of phospholipid synthesis and movement within both mitochondrial membranes. Such remodeling of mitochondrial phospholipid dynamics progresses with the chronological age of a yeast cell and ultimately causes significant changes in mitochondrial membrane lipidome. These changes in the composition of membrane phospholipids alter mitochondrial abundance and morphology, thereby triggering changes in the age-related chronology of such longevity-defining redox processes as mitochondrial respiration, the maintenance of mitochondrial membrane potential, the preservation of cellular homeostasis of mitochondrially produced reactive oxygen species, and the coupling of electron transport to ATP synthesis.

Visualization and analysis of lipopolysaccharide distribution in binary phospholipid bilayers

Energy Technology Data Exchange (ETDEWEB)

Henning, Maria Florencia [Instituto de Investigaciones Bioquimicas La Plata (INIBIOLP), CCT-La Plata, CONICET, Facultad de Ciencias Medicas, UNLP, Calles 60 y 120, 1900 La Plata (Argentina); Sanchez, Susana [Laboratory for Fluorescence Dynamics, University of California-Irvine, Irvine, CA (United States); Bakas, Laura, E-mail: lbakas@biol.unlp.edu.ar [Instituto de Investigaciones Bioquimicas La Plata (INIBIOLP), CCT-La Plata, CONICET, Facultad de Ciencias Medicas, UNLP, Calles 60 y 120, 1900 La Plata (Argentina); Departamento de Ciencias Biologicas, Facultad de Ciencias Exactas, UNLP, Calles 47 y 115, 1900 La Plata (Argentina)

2009-05-22

Lipopolysaccharide (LPS) is an endotoxin released from the outer membrane of Gram-negative bacteria during infections. It have been reported that LPS may play a role in the outer membrane of bacteria similar to that of cholesterol in eukaryotic plasma membranes. In this article we compare the effect of introducing LPS or cholesterol in liposomes made of dipalmitoylphosphatidylcholine/dioleoylphosphatidylcholine on the solubilization process by Triton X-100. The results show that liposomes containing LPS or cholesterol are more resistant to solubilization by Triton X-100 than the binary phospholipid mixtures at 4 {sup o}C. The LPS distribution was analyzed on GUVs of DPPC:DOPC using FITC-LPS. Solid and liquid-crystalline domains were visualized labeling the GUVs with LAURDAN and GP images were acquired using a two-photon microscope. The images show a selective distribution of LPS in gel domains. Our results support the hypothesis that LPS could aggregate and concentrate selectively in biological membranes providing a mechanism to bring together several components of the LPS-sensing machinery.

Investigation of the chemical mechanisms involved in the electropulsation of membranes at the molecular level.

Science.gov (United States)

Breton, Marie; Mir, Lluis M

2018-02-01

The chemical consequences of electropulsation on giant unilamellar vesicles (GUVs), in particular the possible oxidation of unsaturated phospholipids, have been investigated by mass spectrometry, flow cytometry and absorbance methods. Pulse application induced oxidation of the GUV phospholipids and the oxidation level depended on the duration of the pulse. Light and O 2 increased the level of pulse-induced lipid peroxidation whereas the presence of antioxidants either in the membrane or in the solution completely suppressed peroxidation. Importantly, pulse application did not create additional reactive oxygen species (ROS) in GUV-free solution. Lipid peroxidation seems to result from a facilitation of the lipid peroxidation by the ROS already present in the solution before pulsing, not from a direct pulse-induced peroxidation. The pulse would facilitate the entrance of ROS in the core of the membrane, allowing the contact between ROS and lipid chains and provoking the oxidation. Our findings demonstrate that the application of electric pulses on cells could induce the oxidation of the membrane phospholipids since cell membranes contain unsaturated lipids. The chemical consequences of electropulsation will therefore have to be taken into account in future biomedical applications of electropulsation since oxidized phospholipids play a key role in many signaling pathways and diseases. Copyright © 2017 Elsevier B.V. All rights reserved.

Automated thin-film analyses of anhydrous interplanetary dust particles in the analytical electron microscope

Science.gov (United States)

Bradley, J. P.; Germani, M. S.; Brownlee, D. E.

1989-01-01

An AEM apparatus equipped with digital beam control has obtained quantitative point-count analyses of thin sections taken from eight anhydrous chondritic interplanetary dust particles (IDPs); between 200 and 500 X-ray analyses were collected from each thin section and analyzed for Mg, Al, Si, S, Ca, Cr, Mn, Fe, and Ni. Two types of anhydrous chondritic aggregates were observed in the eight IDPs: one highly porous, the other less so. The eight anhydrous IDPs are characterizable as mixtures of fine- and coarse-grained aggregates, large mineral grains, glass, and carbonaceous materials. Their elemental concentrations follow those of solar abundances, suggesting that they are unperturbed by aqueous alteration.

Phospholipid mediated activation of calcium dependent protein kinase 1 (CaCDPK1 from chickpea: a new paradigm of regulation.

Directory of Open Access Journals (Sweden)

Ajay Kumar Dixit

Full Text Available Phospholipids, the major structural components of membranes, can also have functions in regulating signaling pathways in plants under biotic and abiotic stress. The effects of adding phospholipids on the activity of stress-induced calcium dependent protein kinase (CaCDPK1 from chickpea are reported here. Both autophosphorylation as well as phosphorylation of the added substrate were enhanced specifically by phosphatidylcholine and to a lesser extent by phosphatidic acid, but not by phosphatidylethanolamine. Diacylgylerol, the neutral lipid known to activate mammalian PKC, stimulated CaCDPK1 but at higher concentrations. Increase in V(max of the enzyme activity by these phospholipids significantly decreased the K(m indicating that phospholipids enhance the affinity towards its substrate. In the absence of calcium, addition of phospholipids had no effect on the negligible activity of the enzyme. Intrinsic fluorescence intensity of the CaCDPK1 protein was quenched on adding PA and PC. Higher binding affinity was found with PC (K(½ = 114 nM compared to PA (K(½ = 335 nM. We also found that the concentration of PA increased in chickpea plants under salt stress. The stimulation by PA and PC suggests regulation of CaCDPK1 by these phospholipids during stress response.

Synthesis of fertilizers nitrogen and 15N-enriched. Pt. I. Production of enriched 15N-anhydrous ammonia

International Nuclear Information System (INIS)

Bendassolli, J.A.; Mortatti, J.; Trivelin, P.C.O.; Victoria, R.L.

1988-01-01

The results of 15 N-anhydrous ammonia production through reaction between 15 N-enriched ammonium sulphate and sodium hidroxide are reported. Influence of the reaction temperature, carrier gas flow, reaction time and mass of ammonium sulphate on the production of anhydrous ammonia were studied. Analyses for the cost of production of 5% atoms in 15 N-enriched anhydrous ammonia were made. (M.A.C.) [pt

H3PO4 imbibed polyacrylamide-graft-chitosan frameworks for high-temperature proton exchange membranes

Science.gov (United States)

Yuan, Shuangshuang; Tang, Qunwei; He, Benlin; Chen, Haiyan; Li, Qinghua; Ma, Chunqing; Jin, Suyue; Liu, Zhichao

2014-03-01

Proton exchange membrane (PEM), transferring protons from anode to cathode, is a key component in a PEM fuel cell. In the current work, a new class of PEMs are synthesized benefiting from the imbibition behavior of three-dimensional (3D) polyacrylamide-graft-chitosan (PAAm-graft-chitosan) frameworks to H3PO4 aqueous solution. Interconnected 3D framework of PAAm-graft-chitosan provides tremendous space for holding proton-conducting H3PO4. The highest anhydrous proton conductivity of 0.13 S cm-1 at 165 °C is obtained. A fuel cell using a thick membrane as a PEM showed a peak power density of 405 mW cm-2 with O2 and H2 as the oxidant and fuel, respectively. Results indicate that the interconnected 3D framework provides superhighway for proton conduction. The valued merits on anhydrous proton conductivity, huge H3PO4 loading, and easy synthesis promise the new membranes to be good alternatives as high-temperature PEMs.

Effect of docosahexaenoic acid and ascorbate on peroxidation of retinal membranes of ODS rats.

Science.gov (United States)

Wang, Jin-Ye; Sekine, Seiji; Saito, Morio

2003-04-01

Mutant male osteogenic disorder Shionogi (ODS) rats, unable to synthesize ascorbic acid, were fed diets containing a high content of docosahexaenoic acid (DHA) and different amounts of ascorbic acid, to study the effect of DHA on peroxidative susceptibility of the retina and possible antioxidant action of ascorbic acid. ODS rats were fed from 7 weeks of age with diets containing high DHA (6.4% of total energy). A control group received a diet high in linoleic acid. The diets also contained varying amounts of ascorbic acid. Fatty acid compositions and phospholipid hydroperoxides in rod outer segment (ROS) membranes, and retinal ascorbic acid were analyzed. DHA in ROS membranes was significantly increased in rats fed high DHA, compared with the linoleic acid diet. Levels of phospholipid hydroperoxides in the DHA-fed rats were significantly higher than the linoleic acid-fed rats. Ascorbic acid supplementation did not suppress the phospholipid hydroperoxide levels after a high DHA diet, even when the supplement increased the content of retinal ascorbic acid. In conclusion, high DHA feeding induced a marked increase of phospholipid hydroperoxides in ROS membranes of ODS rats. Supplementation of ascorbic acid did not reverse this increase.

Interference of a short-chain phospholipid with ion transport pathways in frog skin

DEFF Research Database (Denmark)

Unmack, M A; Frederiksen, O; Willumsen, N J

1997-01-01

The effects of mucosal application of the short-chain phospholipid didecanoyl-L-alpha-phosphatidylcholine (DDPC; with two saturated 10-carbon acyl chains) on active Na+ transport and transepithelial conductance (G) in the frog skin (Rana temporaria) were investigated. Active Na+ transport...... of the frog skin epithelium and opens a paracellular tight junction pathway. Both effects may be caused by incorporation of DDPC in the apical cell membrane....

Lean hydrous and anhydrous bioethanol combustion in spark ignition engine at idle

International Nuclear Information System (INIS)

Chuepeng, Sathaporn; Srisuwan, Sudecha; Tongroon, Manida

2016-01-01

Highlights: • Anhydrous ethanol burns fastest in uncalibrated engine at equal equivalence ratio. • The leaner hydrous ethanol combustion tends to elevate the COV in imep. • Hydrous ethanol consumption was 10% greater than anhydrous ethanol at ϕ = 0.67 limit. • Optimizing alternative fuel engine at idle for stability and emission is suggested. - Abstract: The applications of anhydrous bioethanol to substitute or replace gasoline fuel have shown to attain benefits in terms of engine thermal efficiency, power output and exhaust emissions from spark ignition engines. A hydrous bioethanol has also been gained more attention due to its energy and cost effectiveness. The main aim of this work is to minimize fuel quantity injected to the intake ports of a four-cylinder engine under idle condition. The engine running with hydrous ethanol undergoes within lean-burn condition as its combustion stability is analyzed using an engine indicating system. Coefficient of variation in indicated mean effective pressure is an indicator for combustion stability with hydrocarbon and carbon monoxide emission monitoring as a supplement. Anhydrous ethanol burns faster than hydrous ethanol and gasoline in the uncalibrated engine at the same fuel-to-air equivalence ratio under idle condition. The leaner hydrous ethanol combustion tends to elevate the coefficient of variation in indicated mean effective pressure. The experimental results have found that the engine consumes greater hydrous ethanol by 10% on mass basis compared with those of anhydrous ethanol at the lean limit of fuel-to-air equivalence ratio of 0.67. The results of exhaust gas analysis were compared with those predicted by chemical equilibrium analysis of the fuel-air combustion; the resemble trends were found. Calibrating the alternative fueled engine for fuel injection quantity should be accomplished at idle with combustion stability and emissions optimization.

Sensing Phosphatidylserine in Cellular Membranes

Directory of Open Access Journals (Sweden)

Jason G. Kay

2011-01-01

Full Text Available Phosphatidylserine, a phospholipid with a negatively charged head-group, is an important constituent of eukaryotic cellular membranes. On the plasma membrane, rather than being evenly distributed, phosphatidylserine is found preferentially in the inner leaflet. Disruption of this asymmetry, leading to the appearance of phosphatidylserine on the surface of the cell, is known to play a central role in both apoptosis and blood clotting. Despite its importance, comparatively little is known about phosphatidylserine in cells: its precise subcellular localization, transmembrane topology and intracellular dynamics are poorly characterized. The recent development of new, genetically-encoded probes able to detect phosphatidylserine within live cells, however, is leading to a more in-depth understanding of the biology of this phospholipid. This review aims to give an overview of the current methods for phosphatidylserine detection within cells, and some of the recent realizations derived from their use.

Phospholipids in Milk Fat: Composition, Biological and Technological Significance, and Analytical Strategies

Directory of Open Access Journals (Sweden)

Giovanna Contarini

2013-01-01

Full Text Available Glycerophospholipids and sphingolipids are quantitatively the most important phospholipids (PLs in milk. They are located on the milk fat globule membrane (MFGM and in other membranous material of the skim milk phase. They include principally phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol and phosphatidylserine, while sphingomyelin is the dominant species of sphingolipids There is considerable evidence that PLs have beneficial health effects, such as regulation of the inflammatory reactions, chemopreventive and chemotherapeutic activity on some types of cancer, and inhibition of the cholesterol absorption. PLs show good emulsifying properties and can be used as a delivery system for liposoluble constituents. Due to the amphiphilic characteristics of these molecules, their extraction, separation and detection are critical points in the analytical approach. The extraction by using chloroform and methanol, followed by the determination by high pressure liquid chromatography (HPLC, coupled with evaporative light scattering (ELSD or mass detector (MS, are the most applied procedures for the PL evaluation. More recently, nuclear magnetic resonance spectrometry (NMR was also used, but despite it demonstrating high sensitivity, it requires more studies to obtain accurate results. This review is focused on milk fat phospholipids; their composition, biological activity, technological properties, and significance in the structure of milk fat. Different analytical methodologies are also discussed.

Interaction of Ketotifen Fumarate with Anhydrous Theophylline in ...

African Journals Online (AJOL)

Purpose: The purpose of the present study was to investigate interaction between ketotifen fumarate and anhydrous theophylline in aqueous media of various pH. Methods: By using Job's continuous-variation analysis and Ardon's spectrophotomeric methods, the values of stability constants of theophylline with ketotifen ...

Structural basis of control of inward rectifier Kir2 channel gating by bulk anionic phospholipids.

Science.gov (United States)

Lee, Sun-Joo; Ren, Feifei; Zangerl-Plessl, Eva-Maria; Heyman, Sarah; Stary-Weinzinger, Anna; Yuan, Peng; Nichols, Colin G

2016-09-01

Inward rectifier potassium (Kir) channel activity is controlled by plasma membrane lipids. Phosphatidylinositol-4,5-bisphosphate (PIP2) binding to a primary site is required for opening of classic inward rectifier Kir2.1 and Kir2.2 channels, but interaction of bulk anionic phospholipid (PL(-)) with a distinct second site is required for high PIP2 sensitivity. Here we show that introduction of a lipid-partitioning tryptophan at the second site (K62W) generates high PIP2 sensitivity, even in the absence of PL(-) Furthermore, high-resolution x-ray crystal structures of Kir2.2[K62W], with or without added PIP2 (2.8- and 2.0-Å resolution, respectively), reveal tight tethering of the C-terminal domain (CTD) to the transmembrane domain (TMD) in each condition. Our results suggest a refined model for phospholipid gating in which PL(-) binding at the second site pulls the CTD toward the membrane, inducing the formation of the high-affinity primary PIP2 site and explaining the positive allostery between PL(-) binding and PIP2 sensitivity. © 2016 Lee et al.

Free and membrane-bound calcium in microgravity and microgravity effects at the membrane level

Science.gov (United States)

Belyavskaya, N. A.

The changes of [Ca^2+]_i controlled is known to play a key regulatory role in numerous cellular processes especially associated with membranes. Previous studies from our laboratory have demonstrated an increase in calcium level in root cells of pea seedlings grown aboard orbital station ``Salyut 6'' /1/. These results: 1) indicate that observed Ca^2+-binding sites of membranes also consist in proteins and phospholipids; 2) suggest that such effects of space flight in membrane Ca-binding might be due to the enhancement of Ca^2+ influx through membranes. In model presented, I propose that Ca^2+-activated channels in plasma membrane in response to microgravity allow the movement of Ca^2+ into the root cells, causing a rise in cytoplasmic free Ca^2+ levels. The latter, in its turn, may induce the inhibition of a Ca^2+ efflux by Ca^2+-activated ATPases and through a Ca^2+/H^+ antiport. It is possible that increased cytosolic levels of Ca^2+ ions have stimulated hydrolysis and turnover of phosphatidylinositols, with a consequent elevation of cytosolic [Ca^2+]_i. Plant cell can response to such a Ca^2+ rise by an enhancement of membranous Ca^2+-binding activities to rescue thus a cell from an abundance of a cytotoxin. A Ca^2+-induced phase separation of membranous lipids assists to appear the structure nonstable zones with high energy level at the boundary of microdomains which are rich by some phospholipid components; there is mixing of molecules of the membranes contacted in these zones, the first stage of membranous fusion, which was found in plants exposed to microgravity. These results support the hypothesis that a target for microgravity effect is the flux mechanism of Ca^2+ to plant cell.

Effects of phosphonium-based ionic liquids on phospholipid membranes studied by small-angle X-ray scattering

Czech Academy of Sciences Publication Activity Database

Kontro, I.; Svedström, K.; Duša, Filip; Ahvenainen, P.; Ruokonen, S. K.; Witos, J.; Wiedmer, S. K.

2016-01-01

Roč. 201, DEC (2016), s. 59-66 ISSN 0009-3084 Institutional support: RVO:68081715 Keywords : phospholipids * x-ray scattering Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 3.361, year: 2016

Lateral interactions in the photoreceptor membrane: a NMR study

International Nuclear Information System (INIS)

Mollevanger, L.C.P.J.

1987-01-01

The photoreceptor membrane has an exceptionally high content of polyunsaturated fatty acyl chains combined with a high amount of phosphatidyl ethanolamine. It is situated in a cell organelle, the rod outer segment, with a high biological activity in which controlable trans-membrane currents of different ions play an important role. These characteristics make it a very interesting biological membrane to search for the existence of non-bilayer structures. Therefore in this thesis a detailed study of the polymorphic phase behaviour of the rod outer segment photoreceptor lipids was undertaken, concerning modulation of the polymorphic phase behaviour of photoreceptor membrane lipids by divalent cations and temperature, polymorphism of the individual phospholipid classes phosphatidylethanolamine and phosphatidylserine and effects of cholesterol, bilayer stabilization by (rhod)opsin. Morphologically intact rod outer segment possesses a large magnetic anisotropy. This property is used to obtain 31 P-NMR of oriented photoreceptor membranes which allows spectral analysis and identification of individual phospholipid classes, and allows to study lateral lipid diffusion in intact disk membranes. The power of high resolution solid state 13 C-NMR to study the conformation of the chromophore in rhodopsin is demonstrated. (Auth.)

Membrane properties involved in calcium-stimulated microparticle release from the plasma membranes of S49 lymphoma cells.

Science.gov (United States)

Campbell, Lauryl E; Nelson, Jennifer; Gibbons, Elizabeth; Judd, Allan M; Bell, John D

2014-01-01

This study answered the question of whether biophysical mechanisms for microparticle shedding discovered in platelets and erythrocytes also apply to nucleated cells: cytoskeletal disruption, potassium efflux, transbilayer phospholipid migration, and membrane disordering. The calcium ionophore, ionomycin, disrupted the actin cytoskeleton of S49 lymphoma cells and produced rapid release of microparticles. This release was significantly inhibited by interventions that impaired calcium-activated potassium current. Microparticle release was also greatly reduced in a lymphocyte cell line deficient in the expression of scramblase, the enzyme responsible for calcium-stimulated dismantling of the normal phospholipid transbilayer asymmetry. Rescue of the scrambling function at high ionophore concentration also resulted in enhanced particle shedding. The effect of membrane physical properties was addressed by varying the experimental temperature (32-42°C). A significant positive trend in the rate of microparticle release as a function of temperature was observed. Fluorescence experiments with trimethylammonium diphenylhexatriene and Patman revealed significant decrease in the level of apparent membrane order along that temperature range. These results demonstrated that biophysical mechanisms involved in microparticle release from platelets and erythrocytes apply also to lymphocytes.

Relationship between red cell membrane fatty acids and adipokines in individuals with varying insulin sensitivity.

Science.gov (United States)

Min, Y; Lowy, C; Islam, S; Khan, F S; Swaminathan, R

2011-06-01

Plasma leptin and adiponectin, and membrane phospholipid fatty acid composition are implicated into the mechanism of insulin resistance but no clear pattern has emerged. Hence, this study examined these variables in subjects presenting to the diabetic clinic for a diagnostic glucose tolerance test. Body composition, glucose, glycated hemoglobin, insulin, leptin, adiponectin, and red cell and plasma phospholipid fatty acids were assessed from 42 normal and 28 impaired glucose tolerant subjects. Insulin sensitivity was determined by homeostatic model assessment. The plasma phosphatidylcholine fatty acid composition of the impaired glucose tolerant subjects was similar to that of normal subjects. However, the impaired glucose tolerant subjects had significantly lower linoleic (Pphosphatidylcholine and phosphatidylethanolamine compared with the normal subjects. Moreover, red cell phosphatidylcholine docosahexaenoic acid correlated positively with adiponectin (r=0.290, Pinsulin (r=-0.335, Pinsulin resistance (r=-0.322, Pinsulin level whereas insulin was the only component that predicted the membrane fatty acids. We postulate that membrane phospholipids fatty acids have an indirect role in determining insulin concentration but insulin has a major role in determining membrane fatty acid composition.

Neutron reflectivity study of substrate surface chemistry effects on supported phospholipid bilayer formation on (1120) sapphire.

Energy Technology Data Exchange (ETDEWEB)

Oleson, Timothy A. [University of Wisconsin, Madison; Sahai, Nita [University of Akron; Wesolowski, David J [ORNL; Dura, Joseph A [ORNL; Majkrzak, Charles F [ORNL; Giuffre, Anthony J. [University of Wisconsin, Madison

2012-01-01

Oxide-supported phospholipid bilayers (SPBs) used as biomimetric membranes are significant for a broad range of applications including improvement of biomedical devices and biosensors, and in understanding biomineralization processes and the possible role of mineral surfaces in the evolution of pre-biotic membranes. Continuous-coverage and/or stacjed SPBs retain properties (e.,g. fluidity) more similar to native biological membranes, which is desirable for most applications. Using neutron reflectivity, we examined face coverage and potential stacking of dipalmitoylphosphatidylcholine (DPPC) bilayers on the (1120) face of sapphire (a-Al2O3). Nearly full bilayers were formed at low to neutral pH, when the sapphire surface is positively charged, and at low ionic strength (l=15 mM NaCl). Coverage decreased at higher pH, close to the isoelectric point of sapphire, and also at high I>210mM, or with addition of 2mM Ca2+. The latter two effects are additive, suggesting that Ca2+ mitigates the effect of higher I. These trends agree with previous results for phospholipid adsorption on a-Al2O3 particles determined by adsorption isotherms and on single-crystal (1010) sapphire by atomic force microscopy, suggesting consistency of oxide surface chemistry-dependent effects across experimental techniques.

Packing of ganglioside-phospholipid monolayers

DEFF Research Database (Denmark)

Majewski, J.; Kuhl, T.L.; Kjær, K.

2001-01-01

Using synchrotron grazing-incidence x-ray diffraction (GIXD) and reflectivity, the in-plane and out-of-plane structure of mixed ganglioside-phospholipid monolayers was investigated at the air-water interface. Mixed monolayers of 0, 5, 10, 20, and 100 mol% ganglioside GM, and the phospholipid...... monolayers did not affect hydrocarbon tail packing (fluidization or condensation of the hydrocarbon region). This is in contrast to previous investigations of lipopolymer-lipid mixtures, where the packing structure of phospholipid monolayers was greatly altered by the inclusion of lipids bearing hydrophilic...

Effects of X-irradiation on membranes of tumor cells

International Nuclear Information System (INIS)

Fonck, K.

1982-01-01

The aim of the investigation was to gain more insight into the effect of ionizing radiation on biomembranes, especially the membrane phospholipids. A general outline of the experimental approach is given in the first chapter. The influence of membrane-active agents and hyperthermia on cell survival after irradiation was studied. Phospholipid turnover was followed by measuring the incorporation of radioactive precursors. The second chapter is an introduction to general radiobiology and to phospholipid metabolism. After the presentation of some physico-chemical properties of ionizing radiation, the effects on cells and cellular components are described. In chapters 3 to 6 the experimental part is described. Chapter 3 starts with the determination of the cellular survival of L5178Y lymphoma cells after X-irradiation. In chapter 4 the lipid composition of lymphosarcoma cell nuclei is presented and in chapter 5 studies on the effect of X-irradiation on the incorporation of palmitate and arachidonate into the phospholipids of lymphosarcoma cells are described. Chapter 6 describes experiments in which lymphosarcoma cells isolated from the spleens of tumor-bearing mice were used to study the effect of a low dose of X-rays (5 Gy) on the incorporation of [ 3 H]palmitate and [ 14 C]arachidonate into the lipids of the tumor cells. These fatty acids were rapidly incorporated especially into the phospholipids of the cells. Chapter 7 contains a general discussion on the experimental results. (Auth.)

Anhydrous Ammonia Training Module. Trainer's Package. Participant's Package.

Science.gov (United States)

Beaudin, Bart; And Others

This document contains a trainer's and a participant's package for teaching employees on site safe handling procedures for working with anhydrous ammonia, especially on farms. The trainer's package includes the following: a description of the module; a competency; objectives; suggested instructional aids; a training outline (or lesson plan) for…

Structural role of lipids in mitochondrial and sarcoplasmic reticulum membranes: freeze-fracture electron microscopy studies

Energy Technology Data Exchange (ETDEWEB)

Packer, L; Mehard, C W; Meissner, G; Zahler, W L; Fleischer, S

1974-01-01

The role of phospholipid in the structure of the membranes of beef heart mitochondria and of the sarcoplasmic reticulum membranes from rabbit skeletal muscle has been investigated by freeze-fracture electron microscopy. Progressive removal of membrane phospholipids, by phospholipase A treatment or detergent treatment, or by organic solvent extraction, results in loss of the smooth background seen in membrane fracture faces and decreased ability of membrane to undergo freeze fracture to yield fracture faces. Instead cross-sections of vesicles or particle clusters are observed. Sarcoplasmic reticulum vesicles have a 9 to 1 asymmetry in the distribution of particles between the convex and concave fracture faces. There is also a wide range of particle size distribution in both of these fracture faces with 85-A particles in greatest number. The removal of membrane associated proteins by detergent extraction does not appreciably change the distribution in particle size. Sarcoplasmic reticulum vesicles were dissolved with detergent and reassembled to form membrane vesicles containing mainly one protein (approx. 90%), i.e., the Ca/sup 2 +/ pump protein, and with a ratio of lipid to protein similar to the original membrane. The reconstituted vesicles readily underwent freeze fracture but the asymmetric particle distribution between the fracture faces was no longer observed. The size distribution of particles in the reconstituted membrane, consisting mainly of Ca/sup 2 +/ pump protein, and phospholipid, was similar in heterogeneity to the original sarcoplasmic reticulum membrane. Thus the heterogeneity in particle size could reflect variation in the orientation of the Ca/sup 2 +/ pump protein within the membrane.

Proton-conducting ionic liquid-based proton exchange membrane fuel cell membranes: The key role of ionomer-ionic liquid interaction

Energy Technology Data Exchange (ETDEWEB)

Martinez, Mathieu; Cointeaux, Laure; Iojoiu, Cristina; Lepretre, Jean-Claude; Sanchez, Jean-Yves [LEPMI, UMR 5631, CNRS-INP-UJF, PHELMA-Campus, BP.75, 1130 rue de la Piscine, 38402 Saint-Martin-d' Heres Cedex (France); Molmeret, Yannick; El Kissi, Nadia [Laboratoire de Rheologie, UMR 5520 CNRS-INPG-UJF, ENSHMG, BP 53, 38041 Grenoble (France); Judeinstein, Patrick [Institut de Chimie Moleculaire et des Materiaux d' Orsay (UMR 8182), Batiment 410, Universite Paris-Sud 11, 91405 Orsay Cedex (France)

2010-09-15

The paper deals with the synthesis and characterisation of proton-conducting ionic liquids (PCILs) and their polymer electrolytes obtained by blending modified Nafion membranes with different concentrations of PCILs. The PCILs are obtained by the neutralization of triethylamine with different organic acids. The first part of the paper studies the influence of acidity and acid structure on PCIL thermal and electrochemical performance, while the second part examines membrane conductivity and reveals it to depend more on PCIL structure than on its intrinsic conductivity. At 130 C, conductivities exceeding 10 mS cm{sup -1} were obtained in fully anhydrous conditions. (author)

Antibiotic interaction with phospholipid monolayers

International Nuclear Information System (INIS)

Gambinossi, F.; Mecheri, B.; Caminati, G.; Nocentini, M.; Puggelli, M.; Gabrielli, G.

2002-01-01

We studied the interactions of tetracycline (TC) antibiotic molecules with phospholipid monolayers with the two-fold aim of elucidating the mechanism of action and providing a first step for the realization of bio-mimetic sensors for such drugs by means of the Langmuir-Blodgett technique. We examined spreading monolayers of three phospholipids in the presence of tetracycline in the subphase by means of surface pressure-area and surface potential-area isotherms as a function of bulk pH. We selected phospholipids with hydrophobic chains of the same length but polar head groups differing either in dimensions and protonation equilibria, i.e. dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylethanolamine (DPPE) and dipalmitoylphosphatidic acid (DPPA). The interaction of tetracycline with the three phospholipids was found to be highly dependent on the electric charge of the antibiotic and on the ionization state of the lipid. Significant interactions are established between the negatively charged form of dipalmitoylphosphatidic acid and the zwitterionic form of tetracycline. The drug was found to migrate at the interface where it is adsorbed underneath or/and among the head groups, depending on the surface pressure of the film, whereas penetration through the hydrophobic layer was excluded for all the three phospholipids

Antibiotic interaction with phospholipid monolayers

Energy Technology Data Exchange (ETDEWEB)

Gambinossi, F.; Mecheri, B.; Caminati, G.; Nocentini, M.; Puggelli, M.; Gabrielli, G

2002-12-01

We studied the interactions of tetracycline (TC) antibiotic molecules with phospholipid monolayers with the two-fold aim of elucidating the mechanism of action and providing a first step for the realization of bio-mimetic sensors for such drugs by means of the Langmuir-Blodgett technique. We examined spreading monolayers of three phospholipids in the presence of tetracycline in the subphase by means of surface pressure-area and surface potential-area isotherms as a function of bulk pH. We selected phospholipids with hydrophobic chains of the same length but polar head groups differing either in dimensions and protonation equilibria, i.e. dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylethanolamine (DPPE) and dipalmitoylphosphatidic acid (DPPA). The interaction of tetracycline with the three phospholipids was found to be highly dependent on the electric charge of the antibiotic and on the ionization state of the lipid. Significant interactions are established between the negatively charged form of dipalmitoylphosphatidic acid and the zwitterionic form of tetracycline. The drug was found to migrate at the interface where it is adsorbed underneath or/and among the head groups, depending on the surface pressure of the film, whereas penetration through the hydrophobic layer was excluded for all the three phospholipids.

Harvesting Water from Air: Using Anhydrous Salt with Sunlight

KAUST Repository

Li, Renyuan; Shi, Yusuf; Shi, Le; Alsaedi, Mossab.; Wang, Peng

2018-01-01

Atmospheric water is abundant alternative water resource, equivalent to 6 times of water in all rivers on Earth. This work screens 14 common anhydrous and hydrated salt couples in terms of their physical and chemical stability, water vapor

Glass transition of anhydrous starch by fast scanning calorimetry.

Science.gov (United States)

Monnier, Xavier; Maigret, Jean-Eudes; Lourdin, Denis; Saiter, Allisson

2017-10-01

By means of fast scanning calorimetry, the glass transition of anhydrous amorphous starch has been measured. With a scanning rate of 2000Ks -1 , thermal degradation of starch prior to the glass transition has been inhibited. To certify the glass transition measurement, structural relaxation of the glassy state has been investigated through physical aging as well as the concept of limiting fictive temperature. In both cases, characteristic enthalpy recovery peaks related to the structural relaxation of the glass have been observed. Thermal lag corrections based on the comparison of glass transition temperatures measured by means of differential and fast scanning calorimetry have been proposed. The complementary investigations give an anhydrous amorphous starch glass transition temperature of 312±7°C. This estimation correlates with previous extrapolation performed on hydrated starches. Copyright © 2017 Elsevier Ltd. All rights reserved.

Domain 4 (D4) of Perfringolysin O to Visualize Cholesterol in Cellular Membranes-The Update.

Science.gov (United States)

Maekawa, Masashi

2017-03-03

The cellular membrane of eukaryotes consists of phospholipids, sphingolipids, cholesterol and membrane proteins. Among them, cholesterol is crucial for various cellular events (e.g., signaling, viral/bacterial infection, and membrane trafficking) in addition to its essential role as an ingredient of steroid hormones, vitamin D, and bile acids. From a micro-perspective, at the plasma membrane, recent emerging evidence strongly suggests the existence of lipid nanodomains formed with cholesterol and phospholipids (e.g., sphingomyelin, phosphatidylserine). Thus, it is important to elucidate how cholesterol behaves in membranes and how the behavior of cholesterol is regulated at the molecular level. To elucidate the complexed characteristics of cholesterol in cellular membranes, a couple of useful biosensors that enable us to visualize cholesterol in cellular membranes have been recently developed by utilizing domain 4 (D4) of Perfringolysin O (PFO, theta toxin), a cholesterol-binding toxin. This review highlights the current progress on development of novel cholesterol biosensors that uncover new insights of cholesterol in cellular membranes.

Interaction of arsenic compounds with model phospholipid membranes

International Nuclear Information System (INIS)

Jemiola-Rzeminska, Malgorzata; Rivera, Cecilia; Suwalsky, Mario; Strzalka, Kazimierz

2007-01-01

This study is part of a project aimed at examining the influence of arsenic on biological membranes. By the use of differential scanning calorimetry (DSC) we have followed the thermotropic behavior of multilamellar vesicles prepared from dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylethanolamine (DMPE) upon incorporation of sodium arsenite (AsI), disodium arsenate (AsII), cacodylic acid (AsIII) and disodium methyl arsenate (AsIV). The effectiveness of perturbations exerted by various arsenic compounds on thermotropic phase transition was further analysed in terms of thermodynamic parameters: transition temperature, enthalpy and molar heat capacity, determined for lipid/As systems on the basis of heating and cooling scans. It is found that while it only has a slight influence on the thermotropic properties of DMPC, arsenic is able to significantly modify DMPE model membranes

Process for the production of sodium carbonate anhydrate

NARCIS (Netherlands)

Oosterhof, H.; Van Rosmalen, G.M.; Witkamp, G.J.; De Graauw, J.

2000-01-01

The invention is directed to a process for the production of sodium carbonate-anhydrate having a bulk density of at least 800 kg/m<3>, said process comprising: providing a suspension of solid sodium carbonate and/or solid sodium bicarbonate and/or solid double salts at least comprising one of sodium

Spectroscopic characterization of cell membranes and their constituents of the plant-associated soil bacterium Azospirillum brasilense

Science.gov (United States)

Kamnev, A. A.; Antonyuk, L. P.; Matora, L. Yu.; Serebrennikova, O. B.; Sumaroka, M. V.; Colina, M.; Renou-Gonnord, M.-F.; Ignatov, V. V.

1999-05-01

Structural and compositional features of bacterial membranes and some of their isolated constituents (cell surface lipopolysaccharide, phospholipids) of the plant-growth-promoting diazotrophic rhizobacterium Azospirillum brasilense (wild-type strain Sp245) were characterized using Fourier transform infrared (FTIR) spectroscopy and some other techniques. FTIR spectra of the cell membranes were shown to comprise the main vibration modes of the relevant lipopolysaccharide and protein components which are believed to be involved in associative plant-bacterium interactions, as well as of phospholipid constituents. The role and functions of metal cations in the structural organization and physicochemical properties of bacterial cell membranes are also discussed considering their accumulation in the membranes from the culture medium.

Accumulation of macular xanthophylls in unsaturated membrane domains.

Science.gov (United States)

Wisniewska, Anna; Subczynski, Witold K

2006-05-15

The distribution of macular xanthophylls, lutein and zeaxanthin, between domains formed in membranes made from an equimolar ternary mixture of dioleoylphosphatidylcholine/sphingomyelin/cholesterol, called a raft-forming mixture, was investigated. In these membranes, two domains are formed: the raft domain enriched in saturated lipids and cholesterol (detergent-resistant membranes, DRM), and the bulk domain enriched in unsaturated lipids (detergent-soluble membranes, DSM). These membrane domains have been separated using cold Triton X-100 extraction from membranes containing 1 mol% of either lutein or zeaxanthin. The results indicated that xanthophylls are substantially excluded from DRM and remain concentrated in DSM. Concentrations of xanthophylls in DRM and DSM calculated as the mole ratio of either xanthophyll to phospholipid were 0.005 and 0.03, respectively, and calculated as the mole ratio of either xanthophyll to total lipid (phospholipid + cholesterol) were 0.003 and 0.025, respectively. Thus, xanthophylls are over eight times more concentrated in DSM than in DRM. No significant difference in the distribution of lutein and zeaxanthin was found. It was also demonstrated using saturation-recovery EPR that at 1 mol%, neither lutein nor zeaxanthin affect the formation of membrane domains. The location of xanthophylls in domains formed from unsaturated lipids is ideal if they are to act as a lipid antioxidant, which is the most accepted mechanism through which lutein and zeaxanthin protect the retina from age-related macular diseases.

Transfer of oleic acid between albumin and phospholipid vesicles

International Nuclear Information System (INIS)

Hamilton, J.A.; Cistola, D.P.

1986-01-01

The net transfer of oleic acid between egg phosphatidylcholine unilamellar vesicles and bovine serum albumin has been monitored by 13 C NMR spectroscopy and 90% isotopically substituted [1- 13 C]oleic acid. The carboxyl chemical shifts of oleic acid bound to albumin were different from those for oleic acid in phospholipid vesicles. Therefore, in mixtures of donor particles, the equilibrium distribution of oleic acid was determined from chemical shift and peak intensity data without separation of donor and acceptor particles. In a system containing equal masses of albumin and phospholipid and a stoichiometry of 4-5 mol of oleic acid per mol of albumin, the oleic acid distribution was pH dependent, with ≥80% of the oleic acid associated with albumin at pH 7.4; association was ≥90% at pH 8.0. Decreasing the pH below 7.4 markedly decreased the proportion of fatty acid bound to albumin. The distribution was reversible with pH and was independent of whether vesicles or albumin acted as a donor. These data suggest that pH may strongly influence the partitioning of fatty acid between cellular membranes and albumin. The 13 C NMR method is also advantageous because it provides information about the structural environments of oleic acid bound to albumin or phospholipid, the ionization state of oleic acid in each environment, and the structural integrity of the vesicles. In addition, minimum and maximum limits for the exchange rates of oleic acid among different environments were obtained from the NMR data

Solvent accessible surface area (ASA) of simulated phospholipid membranes

DEFF Research Database (Denmark)

Tuchsen, E.; Jensen, Morten Østergaard; Westh, P.

2003-01-01

The membrane-solvent interface has been investigated through calculations of the solvent accessible surface area (ASA) for simulated membranes of DPPC and POPE. For DPPC at 52 degreesC we found an ASA of 126 +/- 8 Angstrom(2) per lipid molecule, equivalent to twice the projected lateral area......, even the most exposed parts of the PC head-group show average ASAs of less than half of its maximal or 'fully hydrated' value. The average ASA of a simulated POPE membrane was 96 +/- 7 Angstrom(2) per lipid. The smaller value than for DPPC reflects much lower ASA of the ammonium ion, which is partially...... compensated by increased exposure of the ethylene and phosphate moieties. The ASA of the polar moieties Of (PO4, NH3 and COO) constitutes 65% of the total accessible area for POPE, making this interface more polar than that of DPPC. It is suggested that ASA information can be valuable in attempts...

ESR technique for noninvasive way to quantify cyclodextrins effect on cell membranes

International Nuclear Information System (INIS)

Grammenos, A.; Mouithys-Mickalad, A.; Guelluy, P.H.; Lismont, M.; Piel, G.; Hoebeke, M.

2010-01-01

Research highlights: → ESR: a new tool for cyclodextrins study on living cells. → Cholesterol and phospholipid extraction by Rameb in a dose- and time-dependent way. → Extracted phospholipids and cholesterol form stable aggregates. → ESR spectra show that lipid rafts are damaged by Rameb. → Quantification of the cholesterol extraction on cell membranes in a noninvasive way. -- Abstract: A new way to study the action of cyclodextrin was developed to quantify the damage caused on cell membrane and lipid bilayer. The Electron Spin Resonance (ESR) spectroscopy was used to study the action of Randomly methylated-beta-cyclodextrin (Rameb) on living cells (HCT-116). The relative anisotropy observed in ESR spectrum of nitroxide spin probe (5-DSA and cholestane) is directly related to the rotational mobility of the probe, which can be further correlated with the microviscosity. The use of ESR probes clearly shows a close correlation between cholesterol contained in cells and cellular membrane microviscosity. This study also demonstrates the Rameb ability to extract cholesterol and phospholipids in time- and dose-dependent ways. In addition, ESR spectra enabled to establish that cholesterol is extracted from lipid rafts to form stable aggregates. The present work supports that ESR is an easy, reproducible and noninvasive technique to study the effect of cyclodextrins on cell membranes.

Outcomes of curettage and anhydrous alcohol adjuvant for low-grade chondrosarcoma of long bone.

Science.gov (United States)

Kim, Wanlim; Han, Ilkyu; Kim, Eo Jin; Kang, Seungcheol; Kim, Han-Soo

2015-06-01

Low-grade chondrosarcoma of long bones can be treated successfully with extended intralesional curettage using adjuvants. However, there is no study reporting the use of anhydrous alcohol as an adjuvant in the treatment of low-grade chondrosarcoma. We asked (1) whether intralesional curettage and anhydrous alcohol adjuvant for low-grade chondrosarcoma is associated with good oncologic outcomes; and we report (2) the complications of the procedure. Thirty-six patients (13 men, 23 women) with a mean age of 46 years (range, 18-67 years) were treated for low-grade chondrosarcoma and followed up for a median of 62 months (range, 24-169 months). After intralesional curettage, and additional burring, anhydrous alcohol was used as an adjuvant therapy. At the time of last follow-up, there were no local recurrences or distant metastases. Six patients developed complications: 4 postoperative fractures (11%), 1 intra-articular loose body (3%) and 1 postoperative joint stiffness (3%). Anhydrous alcohol is a reasonable adjuvant for the curettage of low-grade chondrosarcoma of long bones. A long-term follow-up study is necessary, considering the slow biological progression of low-grade chondrosarcoma. Copyright © 2015 Elsevier Ltd. All rights reserved.

Towards Co-evolution of Membranes and Metabolism

Science.gov (United States)

Wei, Chenyu; Wilson, Michael A.; Pohorille, Andrew

2014-01-01

Conceptually, the most robust way to explain how primitive cell-like structures acquired and increased their capabilities is on the basis of Darwinian evolution. A population of protocells containing material that produced more environmentally fit progeny would increase in time at the expense of other protocells. In this scenario, protocellular boundaries were inextricably connected to the metabolism they encapsulated: to be inheritable, early metabolism must have led to an increased rate of growth and division of vesicles and, similarly, transport through vesicle boundaries must have supported the evolution of metabolism. Everything that could not be delivered from the environment had to be produced and retained inside protocells. Despite their importance to the understanding of the origin of life, only a few cases of coupling between metabolism and membrane-related processes have been identified so far. For example, reactions inside fatty-acid vesicles have been linked to their competitive growth and division, and mechanisms by which membrane permeability might have coupled to information polymers have been proposed and explained. Most recently, it has been shown that a dipeptide inside fatty-acid vesicles catalyzes the formation of another dipeptide that binds to vesicle walls and, by doing so, promotes their growth at the expense of other vesicles, thus demonstrating evolutionary advantage of small, membrane-bound peptides. It has been shown that the appearance of phospholipids imparted selective advantage to protocells bound by phospholipid-containing membranes, eventually driving fatty-acid vesicles to extinction. Phospholipid membranes, however, are nearly impermeable to charged species. Yet, the ability to transport ions across membranes was vital for regulating cellular volume, pH homeostasis, generating energy and sensing the environment. To account for this, evolutionary scenarios for the emergence of simple ion channels, protein structures surrounding

The effect of X-irradiation on membrane lipids of lymphosarcoma cells in vivo and in vitro

NARCIS (Netherlands)

Fonck, K; Scherphof, GL; Konings, AWT

1982-01-01

Phospholipids of the membranes of spleen lymphosarcoma cells were radioactively labelled with the radioresistant fatty acid palmitic acid (16:0) and the radiosensitive fatty acid arachidonic acid (20:4). The effect of irradiation on the specific radioactivity of the phospholipids was studied. For

Dietary lipids differentially affect membranes from different areas of rooster sperm.

Science.gov (United States)

Bongalhardo, D C; Leeson, S; Buhr, M M

2009-05-01

The present work aimed to compare the effect of dietary flax with other oil sources on rooster sperm membranes and on semen characteristics. White Leghorn roosters (16 per diet) were fed 1 of 4 treatments: control diet (CON), or a diet containing corn oil (CORN), fish oil (FISH), or flax seed (FLAX) as the lipid source. Semen from 4 birds (30 wk old) of each treatment was pooled, the sperm head (HM) and body membranes (BM) were isolated, and lipids were extracted and analyzed. Aspects of lipid composition tested were as follows: percentage of individual fatty acids (C14:0 to C24:1) in total fatty acids, percentage of fatty acid categories [saturated, monounsaturated, polyunsaturated (PUFA), n-3 and n-6 PUFA, and n-6:n-3 ratio] within total fatty acids, and percentage of phospholipids [phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol, phosphatidylserine, and sphingomyelin] in total phospholipids. Sperm characteristics evaluated were as follows: volume, concentration, viability, percentage of motile cells, average path velocity, track speed, progressive velocity, lateral head displacement, straightness, and linearity. Diet did not affect membrane phospholipid ratios in either membrane but modified major fatty acids within certain phospholipids. Birds fed FISH and CORN showed, respectively, the highest and the lowest n-3 in sperm, causing reciprocal significant changes in n-6:n-3 ratio. Feeding FLAX caused intermediate effects in n-3, with values significantly lower than FISH but higher than CORN in HM (PC, PE, and phosphatidylinositol) and PC in BM (P < 0.05). In the PE phospholipids, FISH, followed by FLAX, increased n-3 in BM and decreased n-6 PUFA in HM. Sperm concentration was specifically correlated with the amount of 20:4n-6 in FLAX and 22:4n-6 in CON. In FLAX diets, straightness correlated with C18:0, n-3, and n-6:n-3 ratio. Diets containing distinct lipid sources differentially modify the lipid contents of HM and BM, with minor

Binding of Neurotransmitters to Lipid Membranes

DEFF Research Database (Denmark)

Peters, Günther H.J.; Werge, Mikkel; Elf-Lind, Maria Northved

2014-01-01

/ acetylated g-aminobutyrate (GABAneu) with a dipalmitoylphosphatidylcholine (DPPC) bilayer. This study was motivated by recent research results that suggested that neural transmission may also be affected by nonspecific interactions of NTs with the lipid matrix of the synaptic membrane. Our results revealed...... backbone of the phospholipids. It is surprising that hydrophilic solutes can deeply penetrate into the membrane pointing to the fact that membrane affinity is governed by specific interactions. Our MD simulations identified the salt-bridge between the primary amine of NTs and the lipid phosphate group...

The role of antioxidant-protein interactions in biological membrane

International Nuclear Information System (INIS)

McGillivray, Duncan J; Singh, Rachna; Melton, Laurence D.; Worcester, David L.; Gilbert, Elliot P.

2009-01-01

Full text: Oxidative damage of cellular membranes has been linked to a variety of disease pathologies, including cardiac disease, Alzheimer's and complications due to diabetes. The oxidation of unsaturated and polyunsaturated fatty acid chains found in cellular membranes leads to significant alteration in membrane physical properties, including lipid orientation and membrane permeability, which ultimately affect biological function. Polyphenols are naturally occurring phytochemicals present in a number of fruit and vegetables that are of interest for their anti-oxidative powers. These polyphenols inhibit lipid oxidation in cellular membrane surfaces, although the mechanism of this inhibition is not entirely clear. Moreover, the polyphenols have significant binding affinity for proteins, which can lead to the formation of soluble and insoluble protein-polyphenol complexes Significantly, in the presence of casein proteins the oxidation inhibition the polyphenols in the membrane is significantly enhanced (as assessed by Lipid Peroxidation Inhibition Capacity assays). Thus the antioxidant pathway appears to involve these protein/polyphenol complexes, as well as direct antioxidant action by the polyphenol. Here we discuss neutron and x-ray scattering results from phospholipid membranes, looking at the positioning of two examples of polyphenolic antioxidants in phospholipid membranes, quercetin and phloretin, the antioxidants' impact on the membrane organisation, and the interaction between antioxidant and extra-membranous protein. This information sheds light on the mechanism of antioxidant protection in these systems, which may be used to understand biological responses to oxidative stress.

Dry fumes purifying system using anhydrous baking soda; Procede chimique d`epuration des fumees au bicarbonate de soude anhydre

Energy Technology Data Exchange (ETDEWEB)

Anon.

1998-04-01

UNISYSTEMS has developed the industrial implementation of the chemical process using anhydrous backing soda, patented by SOLVAY, for purifying fumes containing inorganic salts and sulphur oxides as polluting agents. The system can be applied to industrial processes releasing this type of polluting agents in the fumes at a temperature over 160 deg C, as it is specially indicated in purifying fumes coming from ceramic firing kilns. (authors)

Metformin Decouples Phospholipid Metabolism in Breast Cancer Cells.

Directory of Open Access Journals (Sweden)

Tim A D Smith

Full Text Available The antidiabetic drug metformin, currently undergoing trials for cancer treatment, modulates lipid and glucose metabolism both crucial in phospholipid synthesis. Here the effect of treatment of breast tumour cells with metformin on phosphatidylcholine (PtdCho metabolism which plays a key role in membrane synthesis and intracellular signalling has been examined.MDA-MB-468, BT474 and SKBr3 breast cancer cell lines were treated with metformin and [3H-methyl]choline and [14C(U]glucose incorporation and lipid accumulation determined in the presence and absence of lipase inhibitors. Activities of choline kinase (CK, CTP:phosphocholine cytidylyl transferase (CCT and PtdCho-phospholipase C (PLC were also measured. [3H] Radiolabelled metabolites were determined using thin layer chromatography.Metformin-treated cells exhibited decreased formation of [3H]phosphocholine but increased accumulation of [3H]choline by PtdCho. CK and PLC activities were decreased and CCT activity increased by metformin-treatment. [14C] incorporation into fatty acids was decreased and into glycerol was increased in breast cancer cells treated with metformin incubated with [14C(U]glucose.This is the first study to show that treatment of breast cancer cells with metformin induces profound changes in phospholipid metabolism.

The interaction of insulin with phospholipids

Science.gov (United States)

Perry, M. C.; Tampion, W.; Lucy, J. A.

1971-01-01

1. A simple two-phase chloroform–aqueous buffer system was used to investigate the interaction of insulin with phospholipids and other amphipathic substances. 2. The distribution of 125I-labelled insulin in this system was determined after incubation at 37°C. Phosphatidic acid, dicetylphosphoric acid and, to a lesser extent, phosphatidylcholine and cetyltrimethylammonium bromide solubilized 125I-labelled insulin in the chloroform phase, indicating the formation of chloroform-soluble insulin–phospholipid or insulin–amphipath complexes. Phosphatidylethanolamine, sphingomyelin, cholesterol, stearylamine and Triton X-100 were without effect. 3. Formation of insulin–phospholipid complex was confirmed by paper chromatography. 4. The two-phase system was adapted to act as a simple functional system with which to investigate possible effects of insulin on the structural and functional properties of phospholipid micelles in chloroform, by using the distribution of [14C]glucose between the two phases as a monitor of phospholipid–insulin interactions. The ability of phospholipids to solubilize [14C]glucose in chloroform increased in the order phosphatidylcholineInsulin decreased the [14C]glucose solubilized by phosphatidylcholine, phosphatidylethanolamine and phosphatidic acid, but not by sphingomyelin. 5. The significance of these results and the molecular requirements for the formation of insulin–phospholipid complexes in chloroform are discussed. PMID:5158903

Effect of phospholipid, detergent and protein-protein interaction on stability and phosphoenzyme isomerization of soluble sarcoplasmic reticulum Ca-ATPase.

Science.gov (United States)

Vilsen, B; Andersen, J P

1987-12-30

The purpose of the present study was to elucidate the separate roles of lipid, detergent and protein-protein interaction for stability and catalytic properties of sarcoplasmic reticulum Ca-ATPase solubilized in the non-ionic detergent octa(ethylene glycol) monododecyl ether (C12E8). The use of large-zone high-performance liquid chromatography permitted us to define the self-association state of Ca-ATPase peptide at various detergent, phospholipid and protein concentrations, and also during enzymatic turnover with ATP. Conditions were established for monomerization of Ca-ATPase in the presence of a high concentration of phospholipid relative to detergent. The lipid-saturated monomeric preparation was relatively resistant to inactivation in the absence of Ca2+, whereas delipidated enzyme in monomeric or in oligomeric form was prone to inactivation. Kinetics of phosphoenzyme turnover were examined in the presence and absence of Mg2+. Dephosphorylation rates were sensitive to Mg2+, irrespective of whether the peptide was present in soluble monomeric form or was membrane-bound. C12E8-solubilized monomer without added phospholipid was, however, characterized by a fast initial phase of dephosphorylation in the absence of Mg2+. This was not observed with monomer saturated with phospholipid or with monomer solubilized in myristoylglycerophosphocholine or deoxycholate. The mechanism underlying this difference was shown to be a C12E8-induced acceleration of conversion of ADP-sensitive phosphoenzyme (E1P) to ADP-insensitive phosphoenzyme (E2P). The phosphoenzyme isomerization rate was also found to be enhanced by low-affinity binding of ATP. This was demonstrated both in membrane-bound and in soluble monomeric Ca-ATPase. Our results indicate that a single peptide chain constitutes the target for modulation of phosphoenzyme turnover by Mg2+ and ATP, and that detergent effects, distinct from those arising from disruption of protein-protein contacts, are the major determinants of

Behavior of 4-Hydroxynonenal in Phospholipid Membranes

Czech Academy of Sciences Publication Activity Database

Vazdar, Mario; Jurkiewicz, Piotr; Hof, Martin; Jungwirth, Pavel; Cwiklik, Lukasz

2012-01-01

Roč. 116, č. 22 (2012), s. 6411-6415 ISSN 1520-6106 R&D Projects: GA ČR GA203/08/0114; GA ČR GAP208/10/0376; GA MŠk LH12001; GA AV ČR GEMEM/09/E006 Institutional research plan: CEZ:AV0Z40550506; CEZ:AV0Z40400503 Keywords : fluorescence spectroscopy * molecular dynamics * membrane s * 4-hydroxynonenal Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.607, year: 2012

Hydrophobic nanoparticles promote lamellar to inverted hexagonal transition in phospholipid mesophases.

Science.gov (United States)

Bulpett, Jennifer M; Snow, Tim; Quignon, Benoit; Beddoes, Charlotte M; Tang, T-Y D; Mann, Stephen; Shebanova, Olga; Pizzey, Claire L; Terrill, Nicholas J; Davis, Sean A; Briscoe, Wuge H

2015-12-07

This study focuses on how the mesophase transition behaviour of the phospholipid dioleoyl phosphatidylethanolamine (DOPE) is altered by the presence of 10 nm hydrophobic and 14 nm hydrophilic silica nanoparticles (NPs) at different concentrations. The lamellar to inverted hexagonal phase transition (Lα-HII) of phospholipids is energetically analogous to the membrane fusion process, therefore understanding the Lα-HII transition with nanoparticulate additives is relevant to how membrane fusion may be affected by these additives, in this case the silica NPs. The overriding observation is that the HII/Lα boundaries in the DOPE p-T phase diagram were shifted by the presence of NPs: the hydrophobic NPs enlarged the HII phase region and thus encouraged the inverted hexagonal (HII) phase to occur at lower temperatures, whilst hydrophilic NPs appeared to stabilise the Lα phase region. This effect was also NP-concentration dependent, with a more pronounced effect for higher concentration of the hydrophobic NPs, but the trend was less clear cut for the hydrophilic NPs. There was no evidence that the NPs were intercalated into the mesophases, and as such it was likely that they might have undergone microphase separation and resided at the mesophase domain boundaries. Whilst the loci and exact roles of the NPs invite further investigation, we tentatively discuss these results in terms of both the surface chemistry of the NPs and the effect of their curvature on the elastic bending energy considerations during the mesophase transition.

Ultrafiltration-based degumming of crude rice bran oil using a polymer membrane

Directory of Open Access Journals (Sweden)

Sehn, G. A.R.

2016-03-01

Full Text Available Membrane technology has been gaining momentum in industrial processes, especially in food technology. It is believed to simplify processes, reduce energy consumption, and eliminate pollutants. The objective was to study the performance of polyvinylidene fluoride (PVDF and polyethersulfone (PES polymeric membranes in the degumming of the miscella of crude rice bran oil by using a bench-scale tangential filtration module. In addition, oil miscella filtration techniques using hexane and anhydrous ethyl alcohol solvents were compared. All membranes showed the retention of phospholipids and high flow rates. However, the best performance was observed using the 50-kDa PVDF membrane in miscella hexane solvent, with a 95.5% retention of the phosphorus concentration (by a factor of 1.4, resulting in a permeate with 29 mg·kg−1 of phosphorus and an average flow rate of 48.1 L·m−2·h−1. This technology can be used as a low-pollution, economical alternative for the de-gumming of crude rice bran oil, being effective in the removal of hydratable and non-hydratable phospholipids, resulting in oils with a low phosphorus content.La tecnología de membrana ha ido ganando impulso en los procesos industriales, especialmente en tecnología de los alimentos. Se piensa que simplifica los procesos, reduce el consumo de energía, y elimina contaminantes. El objetivo fué estudiar el rendimiento de las membranas poliméricas de fluoruro de polivinilo (PVDF y poliétersulfona (PES en el desgomado de miscelas de aceite de salvado de arroz crudo, mediante el uso de un módulo de filtración de escalado tangencial. Además, se compararon las técnicas de filtración de miscelas de aceite, utilizando como disolventes hexano y alcohol etílico anhidro. Todas las membranas mostraron retención de los fosfolípidos y altas tasas de flujo. Sin embargo, se observó un mejor rendimiento usando la membrana de PVDF de 50-kDa con hexano como disolvente, con una retención del 95

Electrospun Phospholipid Fibers as Micro-Encapsulation and Antioxidant Matrices.

Science.gov (United States)

Shekarforoush, Elhamalsadat; Mendes, Ana C; Baj, Vanessa; Beeren, Sophie R; Chronakis, Ioannis S

2017-10-17

Electrospun phospholipid (asolectin) microfibers were investigated as antioxidants and encapsulation matrices for curcumin and vanillin. These phospholipid microfibers exhibited antioxidant properties which increased after the encapsulation of both curcumin and vanillin. The total antioxidant capacity (TAC) and the total phenolic content (TPC) of curcumin/phospholipid and vanillin/phospholipid microfibers remained stable over time at different temperatures (refrigerated, ambient) and pressures (vacuum, ambient). ¹H-NMR confirmed the chemical stability of both encapsulated curcumin and vanillin within phospholipid fibers. Release studies in aqueous media revealed that the phenolic bioactives were released mainly due to swelling of the phospholipid fiber matrix over time. The above studies confirm the efficacy of electrospun phospholipid microfibers as encapsulation and antioxidant systems.

Electrospun Phospholipid Fibers as Micro-Encapsulation and Antioxidant Matrices

Directory of Open Access Journals (Sweden)

Elhamalsadat Shekarforoush

2017-10-01

Full Text Available Electrospun phospholipid (asolectin microfibers were investigated as antioxidants and encapsulation matrices for curcumin and vanillin. These phospholipid microfibers exhibited antioxidant properties which increased after the encapsulation of both curcumin and vanillin. The total antioxidant capacity (TAC and the total phenolic content (TPC of curcumin/phospholipid and vanillin/phospholipid microfibers remained stable over time at different temperatures (refrigerated, ambient and pressures (vacuum, ambient. 1H-NMR confirmed the chemical stability of both encapsulated curcumin and vanillin within phospholipid fibers. Release studies in aqueous media revealed that the phenolic bioactives were released mainly due to swelling of the phospholipid fiber matrix over time. The above studies confirm the efficacy of electrospun phospholipid microfibers as encapsulation and antioxidant systems.

Effect of anhydrous magnesium sulphate fertilizer and cutting ...

African Journals Online (AJOL)

Effect of anhydrous magnesium sulphate fertilizer on Panicum maximum yield and chemical composition of the resultant herbage harvested at 3, 4 and 5 weeks of age was investigated. The treatments were labeled T1 to T6. T1 was fertilized and harvested at 3 weeks cutting interval, T2 was fertilized and harvested at 4 ...

The Role of Hydrogen Bonding on Laminar Burning Velocity of Hydrous and Anhydrous Ethanol Fuel with Small Addition of n-Heptane

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I Made Suarta

2016-01-01

Full Text Available The molecular structure of mixed hydrous and anhydrous ethanol with up to 10% v n-heptane had been studied. The burning velocity was examined in a cylindrical explosion combustion chamber. The result showed that the burning velocity of hydrous ethanol is higher than anhydrous ethanol and n-heptane at stoichiometric, rich, and very rich mixtures. The burning velocity of hydrous ethanol with n-heptane drops drastically compared to the burning velocity of anhydrous ethanol with n-heptane. It is caused by two reasons. Firstly, there was a composition change of azeotropic hydrous ethanol molecules within the mixture of fuel. Secondly, at the same volume the number of ethanol molecules in hydrous ethanol was less than in anhydrous ethanol at the same composition of the n-heptane in the mixture. At the mixture of anhydrous ethanol with n-heptane, the burning velocity decreases proportionally to the addition of the n-heptane composition. The burning velocity is between the velocities of anhydrous ethanol and n-heptane. It shows that the burning velocity of anhydrous ethanol mixed with n-heptane is only influenced by the mixture composition.

Supported Phospholipid Membranes Formation at a Gel Electrode and Transport of Divalent Cations across them

Czech Academy of Sciences Publication Activity Database

Navrátil, Tomáš; Šestáková, Ivana; Mareček, Vladimír

2011-01-01

Roč. 6, č. 1 (2011), s. 6032-6046 ISSN 1452-3981 R&D Projects: GA AV ČR IAA400400806; GA ČR GAP206/11/1638 Institutional research plan: CEZ:AV0Z40400503 Keywords : gel * agar * phospholipids Subject RIV: CG - Electrochemistry Impact factor: 3.729, year: 2011 http://www.electrochemsci.org/list11.htm

Desipramine induces disorder in cholesterol-rich membranes: implications for viral trafficking

Science.gov (United States)

Pakkanen, Kirsi; Salonen, Emppu; Mäkelä, Anna R.; Oker-Blom, Christian; Vattulainen, Ilpo; Vuento, Matti

2009-12-01

In this study, the effect of desipramine (DMI) on phospholipid bilayers and parvoviral entry was elucidated. In atomistic molecular dynamics simulations, DMI was found to introduce disorder in cholesterol-rich phospholipid bilayers. This was manifested by a decrease in the deuterium order parameter SCD as well as an increase in the membrane area. Disordering of the membrane suggested DMI to destabilize cholesterol-rich membrane domains (rafts) in cellular conditions. To relate the raft disrupting ability of DMI with novel biological relevance, we studied the intracellular effect of DMI using canine parvovirus (CPV), a virus known to interact with endosomal membranes and sphingomyelin, as an intracellular probe. DMI was found to cause retention of the virus in intracellular vesicular structures leading to the inhibition of viral proliferation. This implies that DMI has a deleterious effect on the viral traffic. As recycling endosomes and the internal vesicles of multivesicular bodies are known to contain raft components, the effect of desipramine beyond the plasma membrane step could be caused by raft disruption leading to impaired endosomal function and possibly have direct influence on the penetration of the virus through an endosomal membrane.

Desipramine induces disorder in cholesterol-rich membranes: implications for viral trafficking

International Nuclear Information System (INIS)

Pakkanen, Kirsi; Mäkelä, Anna R; Oker-Blom, Christian; Vuento, Matti; Salonen, Emppu; Vattulainen, Ilpo

2009-01-01

In this study, the effect of desipramine (DMI) on phospholipid bilayers and parvoviral entry was elucidated. In atomistic molecular dynamics simulations, DMI was found to introduce disorder in cholesterol-rich phospholipid bilayers. This was manifested by a decrease in the deuterium order parameter S CD as well as an increase in the membrane area. Disordering of the membrane suggested DMI to destabilize cholesterol-rich membrane domains (rafts) in cellular conditions. To relate the raft disrupting ability of DMI with novel biological relevance, we studied the intracellular effect of DMI using canine parvovirus (CPV), a virus known to interact with endosomal membranes and sphingomyelin, as an intracellular probe. DMI was found to cause retention of the virus in intracellular vesicular structures leading to the inhibition of viral proliferation. This implies that DMI has a deleterious effect on the viral traffic. As recycling endosomes and the internal vesicles of multivesicular bodies are known to contain raft components, the effect of desipramine beyond the plasma membrane step could be caused by raft disruption leading to impaired endosomal function and possibly have direct influence on the penetration of the virus through an endosomal membrane

Porphyrin-phospholipid interaction and ring metallation depending on the phospholipid polar head type.

Science.gov (United States)

Ramos, Ana P; Pavani, Christiane; Iamamoto, Yassuko; Zaniquelli, Maria E D

2010-10-01

The interaction between a hydrophobically modified 5,10,15,20-tetrakis(4-N-tetradecyl-pyridyl) porphyrin and three phospholipids: two negatively charged, DMPA (the sodium salt of dimyristoyl-sn-glycero-phosphatidyl acid) and DMPG (the sodium salt of 1,2-dimyristoyl-sn-glycero-3-[phospho-rac-(1-glycerol)]) and a zwitterionic DMPC (dimyristoyl-sn-glycero-phosphatidylcholine), were studied by means of surface pressure isotherms and spectroscopic methods. The interaction results in partial or total metallation of the porphyrin with zinc ions in the presence of negatively charged phospholipids, as attested by UV-vis and luminescence spectroscopy of the transferred films. In the presence of the zwitterionic phospholipid no insertion of zinc ion in the porphyrin ring is detected. These results are relevant for the understanding of photosensitizer-lipid-carrier binding for use in photodynamic therapy. Copyright 2010 Elsevier Inc. All rights reserved.

Antidiabetic phospholipid-nuclear receptor complex reveals the mechanism for phospholipid-driven gene regulation

Energy Technology Data Exchange (ETDEWEB)

Musille, Paul M; Pathak, Manish C; Lauer, Janelle L; Hudson, William H; Griffin, Patrick R; Ortlund, Eric A [Emory-MED; (Scripps)

2013-01-31

The human nuclear receptor liver receptor homolog-1 (LRH-1) has an important role in controlling lipid and cholesterol homeostasis and is a potential target for the treatment of diabetes and hepatic diseases. LRH-1 is known to bind phospholipids, but the role of phospholipids in controlling LRH-1 activation remains highly debated. Here we describe the structure of both apo LRH-1 and LRH-1 in complex with the antidiabetic phospholipid dilauroylphosphatidylcholine (DLPC). Together with hydrogen-deuterium exchange MS and functional data, our studies show that DLPC binding is a dynamic process that alters co-regulator selectivity. We show that the lipid-free receptor undergoes previously unrecognized structural fluctuations, allowing it to interact with widely expressed co-repressors. These observations enhance our understanding of LRH-1 regulation and highlight its importance as a new therapeutic target for controlling diabetes.

[Germ cell membrane lipids in spermatogenesis].

Science.gov (United States)

Wang, Ting; Shi, Xiao; Quan, Song

2016-05-01

Spermatogenesis is a complex developmental process in which a diploid progenitor germ cell transforms into highly specialized spermatozoa. During spermatogenesis, membrane remodeling takes place, and cell membrane permeability and liquidity undergo phase-specific changes, which are all associated with the alteration of membrane lipids. Lipids are important components of the germ cell membrane, whose volume and ratio fluctuate in different phases of spermatogenesis. Abnormal lipid metabolism can cause spermatogenic dysfunction and consequently male infertility. Germ cell membrane lipids are mainly composed of cholesterol, phospholipids and glycolipids, which play critical roles in cell adhesion and signal transduction during spermatogenesis. An insight into the correlation of membrane lipids with spermatogenesis helps us to better understand the mechanisms of spermatogenesis and provide new approaches to the diagnosis and treatment of male infertility.

Preparation of pure anhydrous rare earth chlorides

International Nuclear Information System (INIS)

Bel'kova, N.L.; Slastenova, N.M.; Batyaev, I.M.; Solov'ev, M.A.

1979-01-01

A method has been suggested for obtaining extra-pure anhydrous REE chlorides by chloridizing corresponding oxalates by chlorine in a fluid bed, the chloridizing agents being diluted by an inert gas in a ratio of 2-to-1. The method is applicable to the manufacture of quality chlorides not only of light, but also of heavy REE. Neodymium chloride has an excited life of tau=30 μs, this evidencing the absence of the damping impurities

Post-translational regulation of P2X receptor channels: modulation by phospholipids

Directory of Open Access Journals (Sweden)

Louis-Philippe eBernier

2013-11-01

Full Text Available P2X receptor channels mediate fast excitatory signaling by ATP and play major roles in sensory transduction, neuro-immune communication and inflammatory response. P2X receptors constitute a gene family of calcium-permeable ATP-gated cation channels therefore the regulation of P2X signaling is critical for both membrane potential and intracellular calcium homeostasis. Phosphoinositides (PIPn are anionic signaling phospholipids that act as functional regulators of many types of ion channels. Direct PIPn binding was demonstrated for several ligand- or voltage-gated ion channels, however no generic motif emerged to accurately predict lipid-protein binding sites. This review presents what is currently known about the modulation of the different P2X subtypes by phospholipids and about critical determinants underlying their sensitivity to PIPn levels in the plasma membrane.All functional mammalian P2X subtypes tested, with the notable exception of P2X5, have been shown to be positively modulated by PIPn, i.e. homomeric P2X1, P2X2, P2X3, P2X4, and P2X7, as well as heteromeric P2X1/5 and P2X2/3 receptors. Based on various results reported on the aforementioned subtypes including mutagenesis of the prototypical PIPn-sensitive P2X4 and PIPn-insensitive P2X5 receptor subtypes, an increasing amount of functional, biochemical and structural evidence converges on the modulatory role of a short polybasic domain located in the proximal C-terminus of P2X subunits. This linear motif, semi-conserved in the P2X family, seems necessary and sufficient for encoding direct modulation of ATP-gated channels by PIPn. Furthermore, the physiological impact of the regulation of ionotropic purinergic responses by phospholipids on pain pathways was recently revealed in the context of native crosstalks between phospholipase C-linked metabotropic receptors and P2X receptor channels in DRG sensory neurons and microglia.

Mesoscopic models of biological membranes

DEFF Research Database (Denmark)

Venturoli, M.; Sperotto, Maria Maddalena; Kranenburg, M.

2006-01-01

Phospholipids are the main components of biological membranes and dissolved in water these molecules self-assemble into closed structures, of which bilayers are the most relevant from a biological point of view. Lipid bilayers are often used, both in experimental and by theoretical investigations...... to coarse grain a biological membrane. The conclusion of this comparison is that there can be many valid different strategies, but that the results obtained by the various mesoscopic models are surprisingly consistent. A second objective of this review is to illustrate how mesoscopic models can be used...

Interleaflet Coupling, Pinning, and Leaflet Asymmetry—Major Players in Plasma Membrane Nanodomain Formation

Science.gov (United States)

Fujimoto, Toyoshi; Parmryd, Ingela

2017-01-01

The plasma membrane has a highly asymmetric distribution of lipids and contains dynamic nanodomains many of which are liquid entities surrounded by a second, slightly different, liquid environment. Contributing to the dynamics is a continuous repartitioning of components between the two types of liquids and transient links between lipids and proteins, both to extracellular matrix and cytoplasmic components, that temporarily pin membrane constituents. This make plasma membrane nanodomains exceptionally challenging to study and much of what is known about membrane domains has been deduced from studies on model membranes at equilibrium. However, living cells are by definition not at equilibrium and lipids are distributed asymmetrically with inositol phospholipids, phosphatidylethanolamines and phosphatidylserines confined mostly to the inner leaflet and glyco- and sphingolipids to the outer leaflet. Moreover, each phospholipid group encompasses a wealth of species with different acyl chain combinations whose lateral distribution is heterogeneous. It is becoming increasingly clear that asymmetry and pinning play important roles in plasma membrane nanodomain formation and coupling between the two lipid monolayers. How asymmetry, pinning, and interdigitation contribute to the plasma membrane organization is only beginning to be unraveled and here we discuss their roles and interdependence. PMID:28119914

Administration of phospholipide hepatoprotective drug Phosphogliv in patients with psoriatic arthritis (preliminary results

Directory of Open Access Journals (Sweden)

T. V. Korotaeva

2004-01-01

Full Text Available Considering membrane-reparative properties of a new phospholipid hepatoprotector Phosphogliv (FG its therapeutic efficacy was assessed in 9 pts with psoriatic arthritis (PA accompanied by prominent disturbances of blood rheology. FG was given 0,6 g a day during 3 months. Significant decrease of erythrocyte aggregation resulting in increase of erythrocyte aggregation formation time and diminishment of their hydrodynamic resistance without changes of whole blood general caisson viscosity was achieved. Significant improvement of Richie index, tender joint count and pt assessment was noted. The results prove availability of PG administration in PA therapy and possibility of enlargement PG application area owing to membrane-reparative properties of contained in it polyunsaturated phosphatidilcholin in combination with immunomodulating and anti-inflammatory action of glycyrrhizinic acid.

Effect of fish oil supplementation on the n-3 fatty acid content of red blood cell membranes in preterm infants.

Science.gov (United States)

Carlson, S E; Rhodes, P G; Rao, V S; Goldgar, D E

1987-05-01

Very low birth weight infants demonstrate significant reductions in red blood cell membrane docosahexaenoic acid (DHA, 22:6n-3) following delivery unless fed human milk. The purpose of the present study was to determine if a dietary source of DHA (MaxEPA, R. P. Scherer Corporation, Troy, MI) could prevent the decline in red blood cell phospholipid DHA in very low birth weight infants whose enteral feeding consisted of a preterm formula without DHA. Longitudinal data were obtained on membrane phospholipid DHA in both unsupplemented and MaxEPA-supplemented infants by a combination of thin-layer and gas chromatography. These infants (n = 39) ranged in age from 10 to 53 days at enrollment (0 time). At enrollment, phospholipid DHA and arachidonic acid (20:4n-6) were inversely correlated with age in days. During the study, mean red blood cell phospholipid DHA declined without supplementary DHA as determined by biweekly measurement, but infants supplemented with MaxEPA maintained the same weight percent of phospholipid (phosphatidylethanolamine, phosphatidylcholine, and phosphatidylserine) DHA as at enrollment. The pattern of red blood cell phospholipid fatty acids in supplemented infants was similar to that reported for preterm infants fed human milk.

Lysosomotropic cationic amphiphilic drugs inhibit adipocyte differentiation in 3T3-L1K cells via accumulation in cells and phospholipid membranes, and inhibition of autophagy.

Science.gov (United States)

Kagebeck, Patrik; Nikiforova, Violetta; Brunken, Lars; Easwaranathan, Arrabi; Ruegg, Joelle; Cotgreave, Ian; Munic Kos, Vesna

2018-04-05

Some cationic amphiphilic drugs (CADs) have been individually reported to interfere with the differentiation of immune system cells, such as macrophages and dendritic cells. To investigate the possible generic nature of this process, in this study we aimed to see whether these drugs are capable of interfering with the differentiation of adipocytes. Further, we investigated whether this feature might be connected to the lysosomotropic character of these drugs, and their disturbance of intracellular membrane trafficking rather than to the individual pharmacologic properties of each drug. Thus, for the selected set of compounds consisting of seven structurally and pharmacologically diverse CADs and three non-CAD controls we have measured the impact on differentiation of 3T3-L1K murine preadipocytes to adipocytes. We conclude that CADs indeed inhibit adipocyte differentiation, as shown morphologically, at the level of lipid droplet formation and on the expression of genetic markers of adipocytes. Furthermore, the intensity of this inhibitory effect was found to strongly positively correlate with the extent of drug accumulation in adipocytes, with their affinity for phospholipid membranes, as well as with their ability to induce phospholipidosis and inhibit autophagy. Copyright © 2018 Elsevier B.V. All rights reserved.

Toward Anhydrous Proton Conductivity Based on Imidazole Functionalized Mesoporous Silica/Nafion Composite Membranes

International Nuclear Information System (INIS)

Amiinu, Ibrahim Saana; Li, Wei; Wang, Guangjin; Tu, Zhengkai; Tang, Haolin; Pan, Mu; Zhang, Haining

2015-01-01

Highlights: • Imidazole-functionalized mesoporous silica/Nafion composite is formed. • Electrostatic interaction between ionic clusters leads to enhanced molecular rigidity and T g . • Charge transfer resistance decreases with increase in temperature up to 130 °C. • The composite membrane exhibited considerable stability over 70 h at 130 °C. - Abstract: Although Nafion is regarded as the most preferred electrolyte membrane and often used as a benchmark for comparative evaluation of other electrolyte membranes, its wide spread for commercial PEM fuel cells is limited by the poor electrochemical properties at elevated temperatures and low relative humidity conditions. Herein, sol–gel synthesized mesoporous silica functionalized with a protogenic molecule (imidazole) is introduced into the Nafion matrix via a colloid mediated process. The formation of a stable colloid enables homogeneous dispersion of the silica-imidazole nanoparticles without aggregation. Under non-humidified conditions, the amphoteric and self-dissociative character of the tethered imidazole within the matrix functions as a transporting medium to facilitate proton conductivity. The structural and chemical phases are characterized, and qualitatively evaluated by XRD, TEM, FT-IR, TGA, and DMA. The results show that the average proton conductivity of the composite membrane with the optimal amount of functionalized nanoparticles increases progressively to 1.06 × 10 −2 S cm −1 at 130 °C, corresponding to an activation energy of 6.95 kJ mol −1 under non-humidified conditions. The mechanism governing the dynamics of proton conductivity and structural limitations as a function of temperature is discussed

Packing properties 1-alkanols and alkanes in a phospholipid membrane

DEFF Research Database (Denmark)

Westh, Peter

2006-01-01

We have used vibrating tube densitometry to investigate the packing properties of four alkanes and a homologous series of ten alcohols in fluid-phase membranes of dimyristoyl phosphatidylcholine (DMPC). It was found that the volume change of transferring these compounds from their pure states int...... into the membrane core, which is loosely packed. In this region, they partially occupy interstitial (or free-) volume, which bring about a denser molecular packing and generate a negative contribution to Vm(puremem)....... into the membrane, Vm(puremem), was positive for small (C4-C6) 1-alkanols while it was negative for larger alcohols and all alkanes. The magnitude of Vm(puremem) ranged from about +4 cm3/mol for alcohols with an alkyl chain about half the length of the fatty acids of DMPC, to -10 to -15 cm3/mol for the alkanes...

Ion Transport across Biological Membranes by Carborane-Capped Gold Nanoparticles.

Science.gov (United States)

Grzelczak, Marcin P; Danks, Stephen P; Klipp, Robert C; Belic, Domagoj; Zaulet, Adnana; Kunstmann-Olsen, Casper; Bradley, Dan F; Tsukuda, Tatsuya; Viñas, Clara; Teixidor, Francesc; Abramson, Jonathan J; Brust, Mathias

2017-12-26

Carborane-capped gold nanoparticles (Au/carborane NPs, 2-3 nm) can act as artificial ion transporters across biological membranes. The particles themselves are large hydrophobic anions that have the ability to disperse in aqueous media and to partition over both sides of a phospholipid bilayer membrane. Their presence therefore causes a membrane potential that is determined by the relative concentrations of particles on each side of the membrane according to the Nernst equation. The particles tend to adsorb to both sides of the membrane and can flip across if changes in membrane potential require their repartitioning. Such changes can be made either with a potentiostat in an electrochemical cell or by competition with another partitioning ion, for example, potassium in the presence of its specific transporter valinomycin. Carborane-capped gold nanoparticles have a ligand shell full of voids, which stem from the packing of near spherical ligands on a near spherical metal core. These voids are normally filled with sodium or potassium ions, and the charge is overcompensated by excess electrons in the metal core. The anionic particles are therefore able to take up and release a certain payload of cations and to adjust their net charge accordingly. It is demonstrated by potential-dependent fluorescence spectroscopy that polarized phospholipid membranes of vesicles can be depolarized by ion transport mediated by the particles. It is also shown that the particles act as alkali-ion-specific transporters across free-standing membranes under potentiostatic control. Magnesium ions are not transported.

Method for application of nitrogen-15-labeled anhydrous ammonia to small plots

International Nuclear Information System (INIS)

Sanchez, C.A.; Blackmer, A.M.

1987-01-01

A method is described that permits precise application of anhydrous ammonia in bands to plots of the size often used in 15 N-tracer studies. This method involves placing a stainless-steel capillary tube in the soil where the ammonia is to be banded, attaching this tube to a cylinder of ammonia, and then pulling the tube through the soil with deposition of ammonia as an even band. The procedure has marked advantages over previously described methods because it can be used with mixtures of anhydrous ammonia and nitrification inhibitors and because the soil environment at the point of application is representative of the soil environment found when a conventional applicator is used

Understanding Detergent Effects on Lipid Membranes: A Model Study of Lysolipids

DEFF Research Database (Denmark)

Henriksen, Jonas Rosager; Andresen, Thomas Lars; Feldborg, Lise Nørkjær

2010-01-01

Lysolipids and fatty acids are the natural products formed by the hydrolysis of phospholipids. Lysolipids and fatty acids form micelles in solution and acts as detergents in the presence of lipid membranes. In this study, we investigate the detergent strength of a homologous series of lyso......-chain mismatch between LPC and POPC determines the magnitude of the membrane mechanical perturbation per LPC molecule in the membrane. Finally, the three-stage model describing detergent membrane interaction has been extended by a parameter D-MCI, which governs the membrane curvature stability in the detergent...

Engineering monolayer poration for rapid exfoliation of microbial membranes.

Science.gov (United States)

Pyne, Alice; Pfeil, Marc-Philipp; Bennett, Isabel; Ravi, Jascindra; Iavicoli, Patrizia; Lamarre, Baptiste; Roethke, Anita; Ray, Santanu; Jiang, Haibo; Bella, Angelo; Reisinger, Bernd; Yin, Daniel; Little, Benjamin; Muñoz-García, Juan C; Cerasoli, Eleonora; Judge, Peter J; Faruqui, Nilofar; Calzolai, Luigi; Henrion, Andre; Martyna, Glenn J; Grovenor, Chris R M; Crain, Jason; Hoogenboom, Bart W; Watts, Anthony; Ryadnov, Maxim G

2017-02-01

The spread of bacterial resistance to traditional antibiotics continues to stimulate the search for alternative antimicrobial strategies. All forms of life, from bacteria to humans, are postulated to rely on a fundamental host defense mechanism, which exploits the formation of open pores in microbial phospholipid bilayers. Here we predict that transmembrane poration is not necessary for antimicrobial activity and reveal a distinct poration mechanism that targets the outer leaflet of phospholipid bilayers. Using a combination of molecular-scale and real-time imaging, spectroscopy and spectrometry approaches, we introduce a structural motif with a universal insertion mode in reconstituted membranes and live bacteria. We demonstrate that this motif rapidly assembles into monolayer pits that coalesce during progressive membrane exfoliation, leading to bacterial cell death within minutes. The findings offer a new physical basis for designing effective antibiotics.

Bilayer/cytoskeleton interactions in lipid-symmetric erythrocytes assessed by a photoactivable phospholipid analogue

International Nuclear Information System (INIS)

Pradhan, D.; Schlegel, R.A.; Williamson, P.

1991-01-01

Two mechanisms have been proposed for maintenance of transbilayer phospholipid asymmetry in the erythrocyte plasma membrane, one involving specific interactions between the aminophospholipids of the inner leaflet of the bilayer and the cytoskeleton, particularly spectrin, and the other involving the aminophospholipid translocase. If the former mechanism is correct, then erythrocytes which have lost their asymmetric distribution of phospholipids should display altered bilayer/cytoskeleton interactions. To test this possibility, normal erythrocytes, erythrocytes from patients with chronic myelogenous leukemia or sickle disease, and lipid-symmetric and -asymmetric erythrocyte ghosts were labeled with the radioactive photoactivable analogue of phosphatidylethanolamine, 2-(2-azido-4-nitrobenzoyl)-1-acyl-sn-glycero-3-phospho[ 14 C] ethanolamine ([ 14 C]AzPE), previously shown to label cytoskeletal proteins from the bilayer. The labeling pattern of cytoskeletal proteins in pathologic erythrocytes and lipid-asymmetric erythrocyte ghosts was indistinguishable from normal erythrocytes, indicating that the probe detects no differences in bilayer/cytoskeleton interactions in these cells. In contrast, in lipid-symmetric erythrocyte ghosts, labeling of bands 4.1 and 4.2 and actin, and to a lesser extent ankyrin, by [ 14 C]AzPE was considerably reduced. Significantly, however, labeling of spectrin was unaltered in the lipid-symmetric cells. These results do not support a model in which spectrin is involved in the maintenance of an asymmetric distribution of phospholipids in erythrocytes

Tropical to sub-polar gradient in phospholipid composition suggests adaptive tuning of biological membrane function in drosophilds

Czech Academy of Sciences Publication Activity Database

Slotsbo, S.; Sorensen, J. G.; Holmstrup, M.; Košťál, Vladimír; Kellermann, V.; Overgaard, J.

2016-01-01

Roč. 30, č. 5 (2016), s. 759-768 ISSN 0269-8463 Institutional support: RVO:60077344 Keywords : cold tolerance * Drosophila * phospholipid fatty acid composition Subject RIV: ED - Physiology Impact factor: 5.630, year: 2016 http://onlinelibrary.wiley.com/doi/10.1111/1365-2435.12568/abstract

Quantification of fatty acids as methyl esters and phospholipids in cheese samples after separation of triacylglycerides and phospholipids.

Science.gov (United States)

Hauff, Simone; Vetter, Walter

2009-03-23

Determination of the individual fatty acid composition of neutral- and phospholipids as well as the phospholipid content of dairy food and other foodstuffs are important tasks in life sciences. For these purposes, a method was developed for the separation of lipids (standards of triolein and diacylphosphatidylcholines as well as three cheese samples) by solid-phase extraction using a self-packed column filled with partly deactivated silica. Non-halogenated solvents were used for the elution of the lipid classes. Cyclohexane/ethyl acetate (1:1, v/v) served for the elution of neutral lipids, while polar lipids were eluted with three solvents (ethyl acetate/methanol, methanol, and methanol/water) into one fraction. The separated lipid fractions were transesterified and the individual fatty acids were quantified by using gas chromatography coupled to electron ionization mass spectrometry (GC/EI-MS) in the selected ion monitoring (SIM) mode. The recovery rate for standard phosphatidylcholines was approximately 90% and cross-contamination from neutral lipids was negligible. The method was applied to cheese samples. Quantitative amounts of individual fatty acids in the phospholipid fraction were camembert, cheese. Differences in the fatty acid pattern of neutral and polar lipids were detected. The quantity of the fatty acids determined in the phospholipid fraction was divided by the factor 0.7 in order to convert the fatty acid content into the phospholipid content of the cheese samples. This factor is based on the contribution of 16:0 to dipalmitoylphosphatidylcholine (DPPC). The resulting DPPC equivalents (DPPC(eq)) were found to be representative for the average contribution of fatty acids to all classes of phospholipids in dairy products. Using this approach, the phospholipid content of lipids from mozzarella, camembert, and goat cream cheese was 0.60%, 1.42% and 0.79%, respectively.

Digital holographic microscopy of phase separation in multicomponent lipid membranes

Science.gov (United States)

Farzam Rad, Vahideh; Moradi, Ali-Reza; Darudi, Ahmad; Tayebi, Lobat

2016-12-01

Lateral in-homogeneities in lipid compositions cause microdomains formation and change in the physical properties of biological membranes. With the presence of cholesterol and mixed species of lipids, phospholipid membranes segregate into lateral domains of liquid-ordered and liquid-disordered phases. Coupling of two-dimensional intralayer phase separations and interlayer liquid-crystalline ordering in multicomponent membranes has been previously demonstrated. By the use of digital holographic microscopy (DHMicroscopy), we quantitatively analyzed the volumetric dynamical behavior of such membranes. The specimens are lipid mixtures composed of sphingomyelin, cholesterol, and unsaturated phospholipid, 1,2-dioleoyl-sn-glycero-3-phosphocholine. DHMicroscopy in a transmission mode is an effective tool for quantitative visualization of phase objects. By deriving the associated phase changes, three-dimensional information on the morphology variation of lipid stacks at arbitrary time scales is obtained. Moreover, the thickness distribution of the object at demanded axial planes can be obtained by numerical focusing. Our results show that the volume evolution of lipid domains follows approximately the same universal growth law of previously reported area evolution. However, the thickness of the domains does not alter significantly by time; therefore, the volume evolution is mostly attributed to the changes in area dynamics. These results might be useful in the field of membrane-based functional materials.

Dehydration Behaviour of Borax Pentahydrate to Anhydrous Borax by Multi-Stage Heating in a Fluidized

OpenAIRE

ŞAHİN, Ömer

2002-01-01

In order to optimize the anhydrous borax process in a fluidized bed calcinator, it is absolutely necessary to study the dehydration behaviour of borax pentahydrate. It was found that the basic feature of the dehydration of borax pentahydrate to anhydrous borax is concerned with the conditions employed. The bulk density and sodium borate content of the end product were determined to be a function of number of stages of temperature of the dehydration period in the fluidized bed. It...

Transphosphatidylation and base-exchange in the membranes of clostridium butyricum

International Nuclear Information System (INIS)

Walton, P.A.; Goldfine, H.

1987-01-01

Membrane particles from C. butyricum, incubated with phosphatidylserine (PS) and Triton X-100, produced three labelled phospholipids in addition to phosphatidylethanolamine (PE) formed by decarboxylation. These lipid were also formed when PE or phosphatidylglycerol (PG) was employed as substrate. 2D-TLC of intact lipids and their deacylation products showed that these lipids are phosphatidic acid, cardiolipid (CL), and previously unreported phosphatidyltriton (PT). Triton X-100 acts as both activator and substrate of the reaction. With radiolabel in different portions of the substrate phospholipid we demonstrated that the phosphatidyl moiety was transferred with the loss of the head-group in a phospholipase D-like manner. The acceptor molecule can be water or primary alcohols. Addition of exogenous unlabelled PG resulted in the formation of CL with concomitant decrease in the level of PT formed. Labelled PS, PE, and PG could also be formed upon addition of their corresponding alcoholic head group to incubations containing a labelled phosphatidyl donor and Triton X-100. These results indicate that, in C. butyricum, enzymic steps exists that would allow remodelling of the membrane phospholipids, without requiring de novo biosynthesis

1-Oleoyl-2-acetylglycerol stimulates 5-lipoxygenase activity via a putative (phospho)lipid binding site within the N-terminal C2-like domain.

Science.gov (United States)

Hörnig, Christina; Albert, Dana; Fischer, Lutz; Hörnig, Michael; Rådmark, Olof; Steinhilber, Dieter; Werz, Oliver

2005-07-22

5-Lipoxygenase (5-LO) catalysis is positively regulated by Ca2+ ions and phospholipids that both act via the N-terminal C2-like domain of 5-LO. Previously, we have shown that 1-oleoyl-2-acetylglycerol (OAG) functions as an agonist for human polymorphonuclear leukocytes (PMNL) in stimulating 5-LO product formation. Here we have demonstrated that OAG directly stimulates 5-LO catalysis in vitro. In the absence of Ca2+ (chelated using EDTA), OAG strongly and concentration-dependently stimulated crude 5-LO in 100,000 x g supernatants as well as purified 5-LO enzyme from PMNL. Also, the monoglyceride 1-O-oleyl-rac-glycerol and 1,2-dioctanoyl-sn-glycerol were effective, whereas various phospholipids did not stimulate 5-LO. However, in the presence of Ca2+, OAG caused no stimulation of 5-LO. Also, phospholipids or cellular membranes abolished the effects of OAG. As found previously for Ca2+, OAG renders 5-LO activity resistant against inhibition by glutathione peroxidase activity, and this effect of OAG is reversed by phospholipids. Intriguingly, a 5-LO mutant lacking tryptophan residues (Trp-13, -75, and -102) important for the binding of the 5-LO C2-like domain to phospholipids was not stimulated by OAG. We conclude that OAG directly stimulates 5-LO by acting at a phospholipid binding site located within the C2-like domain.

Electrospun Phospholipid Fibers as Micro-Encapsulation and Antioxidant Matrices

DEFF Research Database (Denmark)

Shekarforoush, Elhamalsadat; Mendes, Ana Carina Loureiro; Baj, Vanessa

2017-01-01

Electrospun phospholipid (asolectin) microfibers were investigated as antioxidants and encapsulation matrices for curcumin and vanillin. These phospholipid microfibers exhibited antioxidant properties which increased after the encapsulation of both curcumin and vanillin. The total antioxidant...... capacity (TAC) and the total phenolic content (TPC) of curcumin/phospholipid and vanillin/phospholipid microfibers remained stable over time at different temperatures (refrigerated, ambient) and pressures (vacuum, ambient). ¹H-NMR confirmed the chemical stability of both encapsulated curcumin and vanillin...

Regional distribution of phospholipids in porcine vitreous humor.

Science.gov (United States)

Schnepf, Abigail; Yappert, Marta Cecilia; Borchman, Douglas

2017-07-01

This project explores the regional phospholipid distribution in porcine vitreous humor, retina, and lens. Matrix-assisted laser desorption mass spectrometry has been used previously to image lipids, proteins, and other metabolites in retinas and lenses. However, the regional composition of phospholipids in vitreous humors is not known. To address this issue, we have applied this mass spectral method to explore the regional phospholipid distribution in porcine vitreous humor both ex-situ and in-vitro. To establish the possible source(s) of phospholipids in the vitreous humor, compositional studies of the lens and retina were also pursued. Due to the overall low levels of phospholipids in vitreous humor, it was necessary to optimize the experimental approaches for ex-situ and in-vitro studies. The sensitivity observed in the spectra of methanol extracts from the lens and retina was higher than that for methanol:chloroform extracts, but the compositional trends were the same. A fourfold improvement in sensitivity was observed in the analysis of vitreous humor extracts obtained with the Bligh and Dyer protocol relative to the other two extraction methods. For ex-situ studies, the 'stamp method' with para-nitroaniline as the matrix was chosen. Throughout the vitreous humor, phosphatidylcholines were the most abundant phospholipids. In-vitro results showed higher relative levels of phospholipids compared to the 'stamp' method. However, more details in the regional phospholipid distribution were provided by the ex-situ approach. Both in-vitro and ex-situ results indicated higher levels of phospholipids in the posterior vitreous region, followed by the anterior and central regions. The posterior region contained more unsaturated species whereas more saturated phospholipids were detected in the anterior region. The observed trends suggest that the phospholipids detected in the posterior vitreous humor migrate from the retina and associated vasculature while those present in

Synthesis of anhydrous K2TiOF4 via a mild hydrothermal method

Science.gov (United States)

Felder, Justin B.; Yeon, Jeongho; zur Loye, Hans-Conrad

2015-10-01

The synthesis of anhydrous K2TiOF4 has been previously attempted by transforming precursor compounds, such as the peroxide (K2Ti(O2)F4), hydrate (K2TiOF4·H2O) and fluoride (K2TiF6). Due to the large structural differences between these precursors and the anhydrous oxyfluorides, however, these preparations have been unsuccessful. Therefore, a direct method of synthesis has been employed to grow single crystals of K2TiOF4 that were characterized by single crystal x-ray diffraction. K2TiOF4 was found to be isostructural with the previously known K2VOF4.

7 CFR 58.715 - Cream, plastic cream and anhydrous milkfat.

Science.gov (United States)

2010-01-01

... Service 1 Quality Specifications for Raw Material § 58.715 Cream, plastic cream and anhydrous milkfat. These food products shall be pasteurized, sweet, have a pleasing and desirable flavor and be free from objectionable flavors, and shall be obtained from milk which complies with the quality requirements as specified...

Lipid membrane partitioning of lysolipids and fatty acids: Effects of membrane phase structure and detergent chain length

DEFF Research Database (Denmark)

Høyrup, Lise Pernille Kristine; Davidsen, Jesper; Jørgensen, Kent

2001-01-01

gel phase and at high temperatures in the disordered fluid phase of the phospholipid membrane vesicles. The long saturated acyl chains of the lysolipids and fatty acids varied from 10 to 16 carbon atoms and all titrations were performed below the critical micellar concentrations (cmc...... of magnitude higher when the saturated acyl chain of the detergents increases by two carbon atoms. The obtained partition coefficients are of importance in relation to a deeper understanding of the interplay between global aqueous and local membrane concentrations of the detergents and the functional influence...

Simulation of Water Transport through a Lipid Membrane

NARCIS (Netherlands)

Marrink, Siewert-Jan; Berendsen, Herman J.C.

1994-01-01

To obtain insight in the process of water permeation through a lipid membrane, we performed molecular dynamics simulations on a phospholipid (DPPC)/water system with atomic detail. Since the actual process of permeation is too slow to be studied directly, we deduced the permeation rate indirectly

Dynamic assembly of MinD into filament bundles modulated by ATP, phospholipids, and MinE

OpenAIRE

Suefuji, Kyoko; Valluzzi, Regina; RayChaudhuri, Debabrata

2002-01-01

Accurate positioning of the division septum at the equator of Escherichia coli cells requires a rapid oscillation of MinD ATPase between the polar halves of the cell membrane, together with the division inhibitor MinC, under MinE control. The mechanism underlying MinD oscillation remains poorly understood. Here, we demonstrate that purified MinD assembles into protein filaments in the presence of ATP. Incubation with phospholipid vesicles further stimulates MinD polymerization. Addition of pu...

Platelet activating factor activity in the phospholipids of bovine spermatozoa

Energy Technology Data Exchange (ETDEWEB)

Parks, J.E.; Hough, S.; Elrod, C. (Cornell Univ., Ithaca, NY (USA))

1990-11-01

Platelet activating factor (PAF) has been detected in sperm from several mammalian species and can affect sperm motility and fertilization. Because bovine sperm contain a high percentage of ether-linked phospholipid precursors required for PAF synthesis, a study was undertaken to determine the PAF activity of bovine sperm phospholipids. Total lipids of washed, ejaculated bull sperm were extracted, and phospholipids were fractionated by thin-layer chromatography. Individual phospholipid fractions were assayed for PAF activity on the basis of (3H)serotonin release from equine platelets. PAF activity was detected in the PAF fraction (1.84 pmol/mumol total phospholipid) and in serine/inositol (PS/PI), choline (CP), and ethanolamine phosphoglyceride (EP) and cardiolipin (CA) fractions. Activity was highest in the CP fraction (8.05 pmol/mumol total phospholipid). Incomplete resolution of PAF and neutral lipids may have contributed to the activity in the PS/PI and CA fractions, respectively. Phospholipids from nonsperm sources did not stimulate serotonin release. Platelet activation by purified PAF and by sperm phospholipid fractions was inhibited by the receptor antagonist SRI 63-675. These results indicate that bovine sperm contain PAF and that other sperm phospholipids, especially CP and EP, which are high in glycerylether components, are capable of receptor-mediated platelet activation.

Influence of membrane composition on its flexibility

International Nuclear Information System (INIS)

Gerbelli, B.B.; Teixeira da Silva, E.R.; Oliveira, C.L.P.; Oliveira, E.A.

2012-01-01

Full text: Lamellar phases and vesicles composed of lipids have been used as model systems to investigate biological process related to cell membrane as well as promising carriers for drugs and gene therapy. The composition of the membrane determines its three dimensional shape and its properties such as rigidity and compressibility which play an important role on membrane fusion, protein adhesion, interactions between proteins, etc. We present systematic study of a lamellar system composed mainly of lecithin which is a biocompatible phospholipid and simusol, which is a mixture of fatty acids that acts as a cosurfactant introducing flexibility to the membrane. Using X ray scattering we determine the lamellar periodicity as a function of the hydration for different formulations of the membrane; ranging from 100 % to 50 % mass fraction of lecithin. The X-ray spectra are fitted using a 4 Gaussian model [1]that allows us to determine the lamellar periodicity and the Caille parameter [2]. The ideal swelling law relating the membrane volume fraction (φ m ) to the lamellar periodicity (D) is given by φ m =δ m /D, where δ m is the thickness membrane, however, when steric interactions are dominant with respect to electrostatic and van der Waals interactions, deviations from this behavior are expected [3]. We present experimental data illustrating the swelling behavior for the membrane compositions and the respective behavior of the hydration limit, membrane Luzzati [4], of the Caille parameter and qualitative interpretation of the interaction forces the systems studying the parameter membrane square amplitude fluctuation[5]. [1] Private communication with Prof. Dr. Cristiano Luis Pinto de Oliveira. [2] Caille A. et all, Acad. Sci. Paris B274 (1972) 891. [3] E. Kurtisovski et all, PRL 98, 258103 (2007). [4] Nagle et all, Curr Opin Struct Biol. 2000 Aug;10(4):474-80. [5] H. I. Petrache. Structure and interactions of fluid phospholipids bilayers measured by high resolution

Influence of membrane composition on its flexibility

Energy Technology Data Exchange (ETDEWEB)

Gerbelli, B.B.; Teixeira da Silva, E.R.; Oliveira, C.L.P.; Oliveira, E.A. [Universidade de Sao Paulo (USP), SP (Brazil)

2012-07-01

Full text: Lamellar phases and vesicles composed of lipids have been used as model systems to investigate biological process related to cell membrane as well as promising carriers for drugs and gene therapy. The composition of the membrane determines its three dimensional shape and its properties such as rigidity and compressibility which play an important role on membrane fusion, protein adhesion, interactions between proteins, etc. We present systematic study of a lamellar system composed mainly of lecithin which is a biocompatible phospholipid and simusol, which is a mixture of fatty acids that acts as a cosurfactant introducing flexibility to the membrane. Using X ray scattering we determine the lamellar periodicity as a function of the hydration for different formulations of the membrane; ranging from 100 % to 50 % mass fraction of lecithin. The X-ray spectra are fitted using a 4 Gaussian model [1]that allows us to determine the lamellar periodicity and the Caille parameter [2]. The ideal swelling law relating the membrane volume fraction ({phi}{sub m}) to the lamellar periodicity (D) is given by {phi}{sub m} ={delta}{sub m}/D, where {delta}{sub m} is the thickness membrane, however, when steric interactions are dominant with respect to electrostatic and van der Waals interactions, deviations from this behavior are expected [3]. We present experimental data illustrating the swelling behavior for the membrane compositions and the respective behavior of the hydration limit, membrane Luzzati [4], of the Caille parameter and qualitative interpretation of the interaction forces the systems studying the parameter membrane square amplitude fluctuation[5]. [1] Private communication with Prof. Dr. Cristiano Luis Pinto de Oliveira. [2] Caille A. et all, Acad. Sci. Paris B274 (1972) 891. [3] E. Kurtisovski et all, PRL 98, 258103 (2007). [4] Nagle et all, Curr Opin Struct Biol. 2000 Aug;10(4):474-80. [5] H. I. Petrache. Structure and interactions of fluid phospholipids

High conductive, long-term durable, anhydrous proton conductive solid-state electrolyte based on a metal-organic framework impregnated with binary ionic liquids: Synthesis, characteristic and effect of anion

Science.gov (United States)

Chen, Hui; Han, Shu-Yan; Liu, Rui-Heng; Chen, Teng-Fei; Bi, Kai-Lun; Liang, Jian-Bo; Deng, Yu-Heng; Wan, Chong-Qing

2018-02-01

Incorporating ionic liquids (abbreviated as ILs) into porous metal-organic framework (MOF) to obtain ILs@MOF nanocomposites is documented as a feasible method to achieve new type of anhydrous proton conductor with high performance. We newly synthesized a series of ILs with different acid counter anions (R-SO3-) and their ILs@MOF hybrid materials, i.e. SA-EIMS@MIL-101, MSA-EIMS@MIL-101 and PTSA-EIMS@MIL-101 (SA = sulfate acid, MSA = methanesulfonate acid, PTSA = p-toluenesulfonate acid, EIMS = 1-(1-ethyl-3-imidazolium)propane-3-sulfonate). Such hybrid materials displayed as anhydrous proton conduction with long-term durability even heated at 150 °C open to air. σ value of SA-EIMS@MIL-101 is up to 1.89 × 10-3 S cm-1, being in the range of the most conductive MOF-based materials. MOF support exhibited favorable proton transport and long-term retention for ILs. Anion volumes of R-SO3- displayed significant effects on the proton conductivity of such hybrid ILs@MOF materials. The smaller the van der Waals volume of R-SO3- is, the higher the conductivity of ILs@MOF is. This work suggests that the combination of a variety of the incorporated ILs and a MOF framework would afford high proton transport and gives an idea to explore the safe, anhydrous, solid-state electrolyte for high temperature proton exchange membrane fuel cell.

Membrane curvature, lipid segregation, and structural transitions for phospholipids under dual-solvent stress

International Nuclear Information System (INIS)

Rand, R.P.; Fuller, N.L.; Gruner, S.M.; Parsegian, V.A.

1990-01-01

Amphiphiles respond both to polar and to nonpolar solvents. In this paper X-ray diffraction and osmotic stress have been used to examine the phase behavior, the structural dimensions, and the work of deforming the monolayer-lined aqueous cavities formed by mixtures of dioleoylphosphatidylethanolamine (DOPE) and dioleoylphosphatidylcholine (DOPC) as a function of the concentration of two solvents, water and tetradecane (td). In the absence of td, most PE/PC mixtures show only lamellar phases in excess water; all of these become single reverse hexagonal (H II ) phases with addition of excess td. The spontaneous radius of curvature R 0 of lipid monolayers, as expressed in these H II phases, is allowed by the relief of hydrocarbon chain stress by td; R 0 increases with the ratio DOPC/DOPE. Single H II phases stressed by limited water or td show several responses. (a) the molecular area is compressed at the polar end of the molecule and expanded at the hydrocarbon ends. (b) For circularly symmetrical water cylinders, the degrees of hydrocarbon chain splaying and polar group compression are different for molecules aligned in different directions around the water cylinder. (c) A pivotal position exists along the length of the phospholipid molecule where little area change occurs as the monolayer is bent to increasing curvatures. (d) By defining R 0 at the pivotal position, the authors find that measured energies are well fit by a quadratic bending energy. (e) For lipid mixtures, enforced deviation of the H II monolayer from R 0 is sufficiently powerful to cause demixing of the phospholipids in a way suggesting that the DOPE/DOPC ratio self-adjusts so that its R 0 matches the amount of td or water available, i.e., that curvature energy is minimized

Developing Nanodiscs as a Tool for Low Resolution Studies of Membrane Proteins

DEFF Research Database (Denmark)

Skar-Gislinge, Nicholas

studies of membrane proteins. So far most of the studies using nanodiscs have been concerning the function of the incorporated membrane protein. However, due to the good control of the size and lipid composition of the nanodisc system, they seem an ideal tool for expanding the use of small angle......Phospholipid nanodiscs are ⇠ 10 nm disc shaped particles consisting of about 150 phospholipids arranged in a central bilayer stabilized by two amphipathic protein ”belts” that wrap around the rim of the bilayer. Because they contain a small bilayer leaflet they can be used as a tool for solution......-assembly process in general, and in particular in relation to incorporation of membrane proteins. This was the aim of work done early in this thesis. Here a detailed model for the small angle x-ray and neutron scattering from the empty nanodisc system was derived and used to describe the nanodisc system with great...

Atomistic Monte Carlo Simulation of Lipid Membranes

Directory of Open Access Journals (Sweden)

Daniel Wüstner

2014-01-01

Full Text Available Biological membranes are complex assemblies of many different molecules of which analysis demands a variety of experimental and computational approaches. In this article, we explain challenges and advantages of atomistic Monte Carlo (MC simulation of lipid membranes. We provide an introduction into the various move sets that are implemented in current MC methods for efficient conformational sampling of lipids and other molecules. In the second part, we demonstrate for a concrete example, how an atomistic local-move set can be implemented for MC simulations of phospholipid monomers and bilayer patches. We use our recently devised chain breakage/closure (CBC local move set in the bond-/torsion angle space with the constant-bond-length approximation (CBLA for the phospholipid dipalmitoylphosphatidylcholine (DPPC. We demonstrate rapid conformational equilibration for a single DPPC molecule, as assessed by calculation of molecular energies and entropies. We also show transition from a crystalline-like to a fluid DPPC bilayer by the CBC local-move MC method, as indicated by the electron density profile, head group orientation, area per lipid, and whole-lipid displacements. We discuss the potential of local-move MC methods in combination with molecular dynamics simulations, for example, for studying multi-component lipid membranes containing cholesterol.

Synaptic Membrane Synthesis in Rats Depends on Dietary Sufficiency of Vitamin C, Vitamin E, and Selenium: Relevance for Alzheimer's Disease.

Science.gov (United States)

Cansev, Mehmet; Turkyilmaz, Mesut; Sijben, John W C; Sevinc, Cansu; Broersen, Laus M; van Wijk, Nick

2017-01-01

Chronic consumption of a diet enriched with nutritional precursors of phospholipids, including uridine and the polyunsaturated fatty acids, docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), was shown previously to enhance levels of brain phospholipids and synaptic proteins in rodents. Vitamin C, vitamin E, and selenium may directly affect the breakdown or synthesis of membrane phospholipids. The present study investigated the necessity of antioxidants for the effectiveness of supplementation with uridine plus DHA and EPA (as fish oil) in rats. Rats were randomized to four treatment groups and received, for 6 weeks, one of four experimental diets, i.e., a diet low in antioxidants, a diet high in antioxidants, a diet low in antioxidants supplemented with DHA+EPA+uridine, or a diet high in antioxidants supplemented with DHA+EPA+uridine. On completion of dietary treatment, rats were sacrificed, and brain levels of phospholipids, synaptic proteins, and two enzymes involved in phospholipid synthesis (choline-phosphate cytidylyltransferase, PCYT1A, and choline/ethanolamine phosphotransferase, CEPT1) were analyzed. Levels of phospholipids, the pre- and post-synaptic proteins Synapsin-1 and PSD95, and the enzymes PCYT1A and CEPT1 were significantly enhanced by combined supplementation of DHA+EPA+uridine and antioxidants and not enhanced by supplementation of DHA+EPA+uridine with insufficient antioxidant levels. Our data suggest that dietary vitamin C, vitamin E, and selenium are essential for the phospholipid precursors' effects on increasing levels of membrane phospholipids and synaptic proteins, the indirect indicators of synaptogenesis. Their concomitant supply may be relevant in Alzheimer's disease patients, because the disease is characterized by synapse loss and lower plasma and brain levels of phospholipid precursors and antioxidants.

Anhydrous crystals of DNA bases are wide gap semiconductors.

Science.gov (United States)

Maia, F F; Freire, V N; Caetano, E W S; Azevedo, D L; Sales, F A M; Albuquerque, E L

2011-05-07

We present the structural, electronic, and optical properties of anhydrous crystals of DNA nucleobases (guanine, adenine, cytosine, and thymine) found after DFT (Density Functional Theory) calculations within the local density approximation, as well as experimental measurements of optical absorption for powders of these crystals. Guanine and cytosine (adenine and thymine) anhydrous crystals are predicted from the DFT simulations to be direct (indirect) band gap semiconductors, with values 2.68 eV and 3.30 eV (2.83 eV and 3.22 eV), respectively, while the experimentally estimated band gaps we have measured are 3.83 eV and 3.84 eV (3.89 eV and 4.07 eV), in the same order. The electronic effective masses we have obtained at band extremes show that, at low temperatures, these crystals behave like wide gap semiconductors for electrons moving along the nucleobases stacking direction, while the hole transport are somewhat limited. Lastly, the calculated electronic dielectric functions of DNA nucleobases crystals in the parallel and perpendicular directions to the stacking planes exhibit a high degree of anisotropy (except cytosine), in agreement with published experimental results.

The N-terminal segment of pulmonary surfactant lipopeptide SP-C has intrinsic propensity to interact with and perturb phospholipid bilayers

DEFF Research Database (Denmark)

Plasencia, Inés; Rivas, Luis; Keough, Kevin M W

2004-01-01

aggregation, and leakage of the aqueous content of the vesicles. The lipid-peptide interaction includes a significant hydrophobic component for both zwitterionic and anionic membranes, although the interaction with phosphatidylglycerol bilayers is also electrostatic in nature. The effects of the SP-C N......-terminal peptides on the membrane structure are mediated by significant perturbations of the packing order and mobility of phospholipid acyl chain segments deep in the bilayer, as detected by differential scanning calorimetry and spin-label ESR. These results suggest that the N-terminal region of SP-C, even...

Quantification of fatty acids as methyl esters and phospholipids in cheese samples after separation of triacylglycerides and phospholipids

Energy Technology Data Exchange (ETDEWEB)

Hauff, Simone [University of Hohenheim, Institute of Food Chemistry, Garbenstrasse 28, D-70599 Stuttgart (Germany); Vetter, Walter [University of Hohenheim, Institute of Food Chemistry, Garbenstrasse 28, D-70599 Stuttgart (Germany)], E-mail: w-vetter@uni-hohenheim.de

2009-03-23

Determination of the individual fatty acid composition of neutral- and phospholipids as well as the phospholipid content of dairy food and other foodstuffs are important tasks in life sciences. For these purposes, a method was developed for the separation of lipids (standards of triolein and diacylphosphatidylcholines as well as three cheese samples) by solid-phase extraction using a self-packed column filled with partly deactivated silica. Non-halogenated solvents were used for the elution of the lipid classes. Cyclohexane/ethyl acetate (1:1, v/v) served for the elution of neutral lipids, while polar lipids were eluted with three solvents (ethyl acetate/methanol, methanol, and methanol/water) into one fraction. The separated lipid fractions were transesterified and the individual fatty acids were quantified by using gas chromatography coupled to electron ionization mass spectrometry (GC/EI-MS) in the selected ion monitoring (SIM) mode. The recovery rate for standard phosphatidylcholines was {approx}90% and cross-contamination from neutral lipids was negligible. The method was applied to cheese samples. Quantitative amounts of individual fatty acids in the phospholipid fraction were <0.002-0.29% of total lipids from camembert, <0.002-0.12% of total lipids from mozzarella, and <0.002-0.18% of total lipids in a goat cream cheese. Differences in the fatty acid pattern of neutral and polar lipids were detected. The quantity of the fatty acids determined in the phospholipid fraction was divided by the factor 0.7 in order to convert the fatty acid content into the phospholipid content of the cheese samples. This factor is based on the contribution of 16:0 to dipalmitoylphosphatidylcholine (DPPC). The resulting DPPC equivalents (DPPC{sub eq}) were found to be representative for the average contribution of fatty acids to all classes of phospholipids in dairy products. Using this approach, the phospholipid content of lipids from mozzarella, camembert, and goat cream cheese

Transferred nuclear Overhauser effect analyses of membrane-bound enkephalin analogues by sup 1 H nuclear magnetic resonance: Correlation between activities and membrane-bound conformations

Energy Technology Data Exchange (ETDEWEB)

Milon, Alain; Miyazawa, Tatsuo; Higashijima, Tsutomu (Univ. of Tokyo (Japan))

1990-01-09

Leu-enkephalin, (D-Ala{sup 2})Leu-enkephalin, and (D-Ala{sup 2})Leu-enkephalinamide (agonists) and (L-Ala{sup 2})Leu-enkephalin (inactive analogue) bind to lipid bilayer consisting of phosphatidylcholine and phosphatidylserine. The conformations that these compounds assume, once bound to perdeuterated phospholipid bilayer, have been shown to be unique, as shown by the transferred nuclear Overhauser effect (TRNOE) of {sup 1}H NMR spectroscopy. In addition, their location in the bilayer was analyzed by TRNOE in the presence of spin-labeled phospholipids. These analyses showed a clear relationship between the activity and the peptide-membrane interaction. The three active peptides, when bound to membranes, adopt the same conformation, characterized by a type II{prime} {beta}-turn around Gly{sup 3}-Phe and a {gamma}-turn around Gly{sup 2} (or D-Ala{sup 2}). The inactive analogue, (L-Ala{sup 2})Leu-enkephalin, displayed a completely different TRNOE pattern corresponding to a different conformation in the membrane-bound state. The tyrosine residue of the active compounds is not inserted into the interior of membrane, but it is inserted into the bilayer for the L-Ala{sup 2} analogue. According to these results, (L-Ala{sup 2})Leu-enkephalin may be explained to be inactive because the mode of binding to the membranes is different from that of active compounds.

Rheological properties of poly(vinyl alcohol) (PVA) derived composite membranes for fuel cells

Science.gov (United States)

Remiš, T.

2017-01-01

Rheological properties of new anhydrous proton conducting membrane based on PVA, tetraethyl orthosilicate (TEOS),sulfosuccinic acid (SSA), titanium dioxide (TiO2)was examined at various stoichiometric ratios. SSA was used as sulfonating agents to form a crosslinked structure and as proton source, whereas TEO Sand TiO2were utilized to improve the thermal and mechanical properties of the membrane. In order to verify that all the substances were immobilized into the matrix, the membranes were analysed by means of FT-IR. The rheological, mechanical and thermal properties of the membranes were investigated using rheometer ARES G2 and thermogravimetic analyser (TGA).The analysis of mixed PVA solutions exhibited a unique behaviour of viscosity with increased crosslink density. The dynamic storage modulus G´ of dried composite membranes shows better mechanical resistance and increased tolerance to pressure applied during membrane electrode assembly (MEA).

Some physical properties of anhydrous and hydrated Brownmillerite doped with NaF

International Nuclear Information System (INIS)

Hassaan, M.Y.; El Desoky, M.M.; Salem, S.M.; Yousif, A.A.

2003-01-01

Different samples of Brownmillerite (the ferrite phase of cement clinker) doped with 0, 1 or 3 wt.% NaF were prepared. At first, the oxide mixture of Brownmillerite was prepared according to the following composition: 4 mol CaO, 1 mol Al 2 O 3 and 1 mol Fe 2 O 3 in addition to 1 or 3 wt.% NaF. Each mixture was mixed very well, introduced into an electric furnace at 1300 deg. C for 1 h in a platinum crucible, and then quenched in air. The product was divided into four portions mixed with 40 wt.% distilled water to form Brownmillerite paste, except for one portion which was left dry. Each paste was molded into two molds; after 24 h, they were immersed in a distilled water and withdrawn after 1 or 3 days of hydration, respectively. The pastes were ground again. The anhydrous powders of Brownmillerites and the hydrated samples were prepared for a.c. conduction measurements by pressing it to be in pellets form. The two surfaces of each pellet were coated with silver paste. The a.c. conductivity and dielectric constant for different samples were measured using four-probe method. The data was collected from 320 up to 670 K. Moessbauer spectra and X-ray diffraction patterns were measured for each sample (anhydrous and hydrated) to confirm the formation of Brownmillerite, identify the iron states and the magnetic properties. The results showed that NaF addition to Brownmillerite expedites the hydration reaction rate. The superparamagnetic relaxation, which appeared in the anhydrous Brownmillerite spectra due to the small particle size, decreases with increasing the hydration time. Also, the Fe 3+ (Oh) state increases while Fe 3+ (Td) decreases with the time of hydration. The a.c. conductivity value at fixed frequency for anhydrous and hydrated samples was found to increase with NaF addition. The a.c. conductivity and Moessbauer measurements can be used as good tools to verify the purity of Brownmillerite phase and, accordingly, the purity of cement

Lipophilic drug transfer between liposomal and biological membranes

DEFF Research Database (Denmark)

Fahr, Alfred; van Hoogevest, Peter; Kuntsche, Judith

2006-01-01

This review presents the current knowledge on the interaction of lipophilic, poorly water soluble drugs with liposomal and biological membranes. The center of attention will be on drugs having the potential to dissolve in a lipid membrane without perturbing them too much. The degree of interaction...... is described as solubility of a drug in phospholipid membranes and the kinetics of transfer of a lipophilic drug between membranes. Finally, the consequences of these two factors on the design of lipid-based carriers for oral, as well as parenteral use, for lipophilic drugs and lead selection of oral...... lipophilic drugs is described. Since liposomes serve as model-membranes for natural membranes, the assessment of lipid solubility and transfer kinetics of lipophilic drug using liposome formulations may additionally have predictive value for bioavailability and biodistribution and the pharmacokinetics...

Acyl transfer from membrane lipids to peptides is a generic process.

Science.gov (United States)

Dods, Robert H; Bechinger, Burkhard; Mosely, Jackie A; Sanderson, John M

2013-11-15

The generality of acyl transfer from phospholipids to membrane-active peptides has been probed using liquid chromatography-mass spectrometry analysis of peptide-lipid mixtures. The peptides examined include melittin, magainin II, PGLa, LAK1, LAK3 and penetratin. Peptides were added to liposomes with membrane lipid compositions ranging from pure phosphatidylcholine (PC) to mixtures of PC with phosphatidylethanolamine, phosphatidylserine or phosphatidylglycerol. Experiments were typically conducted at pH7.4 at modest salt concentrations (90 mM NaCl). In favorable cases, lipidated peptides were further characterized by tandem mass spectrometry methods to determine the sites of acylation. Melittin and magainin II were the most reactive peptides, with significant acyl transfer detected under all conditions and membrane compositions. Both peptides were lipidated at the N-terminus by transfer from PC, phosphatidylethanolamine, phosphatidylserine or phosphatidylglycerol, as well as at internal sites: lysine for melittin; serine and lysine for magainin II. Acyl transfer could be detected within 3h of melittin addition to negatively charged membranes. The other peptides were less reactive, but for each peptide, acylation was found to occur in at least one of the conditions examined. The data demonstrate that acyl transfer is a generic process for peptides bound to membranes composed of diacylglycerophospholipids. Phospholipid membranes cannot therefore be considered as chemically inert toward peptides and by extension proteins. © 2013. Published by Elsevier Ltd. All rights reserved.

Process for the production of sodium carbonate anhydrate

OpenAIRE

Oosterhof, H.; Van Rosmalen, G.M.; Witkamp, G.J.; De Graauw, J.

2000-01-01

The invention is directed to a process for the production of sodium carbonate-anhydrate having a bulk density of at least 800 kg/m, said process comprising: providing a suspension of solid sodium carbonate and/or solid sodium bicarbonate and/or solid double salts at least comprising one of sodium carbonate and sodium bicarbonate, in a mixture containing water and an organic, water miscible or partly water miscible solvent, which solvent influences the transition temperature below which sodium...

An adhesion-based method for plasma membrane isolation: evaluating cholesterol extraction from cells and their membranes.

Science.gov (United States)

Bezrukov, Ludmila; Blank, Paul S; Polozov, Ivan V; Zimmerberg, Joshua

2009-11-15

A method to isolate large quantities of directly accessible plasma membrane from attached cells is presented. The method is based on the adhesion of cells to an adsorbed layer of polylysine on glass plates, followed by hypotonic lysis with ice-cold distilled water and subsequent washing steps. Optimal conditions for coating glass plates and time for cell attachment were established. No additional chemical or mechanical treatments were used. Contamination of the isolated plasma membrane by cell organelles was less than 5%. The method uses inexpensive, commercially available polylysine and reusable glass plates. Plasma membrane preparations can be made in 15 min. Using this method, we determined that methyl-beta-cyclodextrin differentially extracts cholesterol from fibroblast cells and their plasma membranes and that these differences are temperature dependent. Determination of the cholesterol/phospholipid ratio from intact cells does not reflect methyl-beta-cyclodextrin plasma membrane extraction properties.

Mechanisms of Membrane Binding of Small GTPase K-Ras4B Farnesylated Hypervariable Region*

Science.gov (United States)

Jang, Hyunbum; Abraham, Sherwin J.; Chavan, Tanmay S.; Hitchinson, Ben; Khavrutskii, Lyuba; Tarasova, Nadya I.; Nussinov, Ruth; Gaponenko, Vadim

2015-01-01

K-Ras4B belongs to a family of small GTPases that regulates cell growth, differentiation and survival. K-ras is frequently mutated in cancer. K-Ras4B association with the plasma membrane through its farnesylated and positively charged C-terminal hypervariable region (HVR) is critical to its oncogenic function. However, the structural mechanisms of membrane association are not fully understood. Here, using confocal microscopy, surface plasmon resonance, and molecular dynamics simulations, we observed that K-Ras4B can be distributed in rigid and loosely packed membrane domains. Its membrane binding domain interaction with phospholipids is driven by membrane fluidity. The farnesyl group spontaneously inserts into the disordered lipid microdomains, whereas the rigid microdomains restrict the farnesyl group penetration. We speculate that the resulting farnesyl protrusion toward the cell interior allows oligomerization of the K-Ras4B membrane binding domain in rigid microdomains. Unlike other Ras isoforms, K-Ras4B HVR contains a single farnesyl modification and positively charged polylysine sequence. The high positive charge not only modulates specific HVR binding to anionic phospholipids but farnesyl membrane orientation. Phosphorylation of Ser-181 prohibits spontaneous farnesyl membrane insertion. The mechanism illuminates the roles of HVR modifications in K-Ras4B targeting microdomains of the plasma membrane and suggests an additional function for HVR in regulation of Ras signaling. PMID:25713064

Milk phospholipids: Organic milk and milk rich in conjugated linoleic acid compared with conventional milk.

Science.gov (United States)

Ferreiro, T; Gayoso, L; Rodríguez-Otero, J L

2015-01-01

The objective of this study was to compare the phospholipid content of conventional milk with that of organic milk and milk rich in conjugated linoleic acid (CLA). The membrane enclosing the fat globules of milk is composed, in part, of phospholipids, which have properties of interest for the development of so-called functional foods and technologically novel ingredients. They include phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylcholine (PC), phosphatidylserine (PS), and the sphingophospholipid sphingomyelin (SM). Milk from organically managed cows contains higher levels of vitamins, antioxidants, and unsaturated fatty acids than conventionally produced milk, but we know of no study with analogous comparisons of major phospholipid contents. In addition, the use of polyunsaturated-lipid-rich feed supplement (extruded linseed) has been reported to increase the phospholipid content of milk. Because supplementation with linseed and increased unsaturated fatty acid content are the main dietary modifications used for production of CLA-rich milk, we investigated whether these modifications would lead to this milk having higher phospholipid content. We used HPLC with evaporative light scattering detection to determine PE, PI, PC, PS, and SM contents in 16 samples of organic milk and 8 samples of CLA-rich milk, in each case together with matching reference samples of conventionally produced milk taken on the same days and in the same geographical areas as the organic and CLA-rich samples. Compared with conventional milk and milk fat, organic milk and milk fat had significantly higher levels of all the phospholipids studied. This is attributable to the differences between the 2 systems of milk production, among which the most influential are probably differences in diet and physical exercise. The CLA-rich milk fat had significantly higher levels of PI, PS, and PC than conventional milk fat, which is also attributed to dietary differences: rations for

Increased phorbol 12,13-dibutyrate (PDBu) receptor function associated with sickle red cell membrane ghosts

International Nuclear Information System (INIS)

Ramachandran, M.; Nair, C.N.; Abraham, E.C.

1987-01-01

The biological receptor for tumor-promoting phorbol esters has been identified as the Ca 2+ /phospholipid dependent enzyme, protein kinase C. In the red cell, this enzyme is mainly cytosolic but becomes translocated to the membrane if the cellular Ca 2+ is allowed to rise. Since cellular Ca 2+ in sickle red cells is high, it was reasoned that this enzyme may become more membrane-bound. In fact, the authors noticed a four-fold increase in the binding of 3 H-PDBu by membrane ghosts isolated from sickle red cells compared to normal red cells (pmoles PDBu bound/mg protein; normal = 0.3 vs sickle cell = 1.4). Attempts to assay the enzyme directly as phospholipid-activated 32 P incorporation into the acid-precipitable membrane proteins also indicated a two-fold increase in the radiolabelling of sickle cell membrane ghosts. Autophosphorylation of membrane proteins and analysis of the phosphorylation profile by SDS-PAGE and autoradiography revealed phosphorylation predominantly of bands 3, 4.1 and 4.9 which are known protein kinase C substrates for the red cell enzyme. The increased membrane-associated protein kinase C in sickle red cells may have a bearing on the altered membrane properties reported in this condition

Effect of phospholipid deposits on adhesion of bacteria to contact lenses.

Science.gov (United States)

Babaei Omali, Negar; Proschogo, Nicholas; Zhu, Hua; Zhao, Zhenjun; Diec, Jennie; Borazjani, Roya; Willcox, Mark D P

2012-01-01

Protein and lipid deposits on contact lenses may contribute to clinical complications. This study examined the effect of phospholipids on the adhesion of bacteria to contact lenses. Worn balafilcon A (n = 11) and senofilcon A (n = 11) were collected after daily wear and phospholipids were extracted in chloroform:methanol. The amount of phospholipid was measured by electrospray ionization mass spectrometry. Unworn lenses soaked in phospholipids were exposed to Pseudomonas aeruginosa and Staphylococcus aureus. After 18 h incubation, the numbers of P. aeruginosa or S. aureus that adhered to the lenses were measured. Phospholipid was tested for possible effects on bacterial growth. A broad range of sphingomyelins (SM) and phosphatidylcholines (PC) were detected from both types of worn lenses. SM (16:0) (m/z 703) and PC (34:2) (m/z 758) were the major phospholipids detected in the lens extracts. Phospholipids did not alter the adhesion of any strain of P. aeruginosa or S. aureus (p > 0.05). Phospholipids (0.1 mg/mL) showed no effect on the growth of P. aeruginosa 6294 or S. aureus 031. Phospholipids adsorb/absorb to contact lenses during wear, however, the major types of phospholipids adsorbed to lenses do not alter bacterial adhesion or growth.

Modulation and interactions of charged biomimetic membranes with bivalent ions

Science.gov (United States)

Kazadi Badiambile, Adolphe

The biological membrane of an eukaryotic cell is a two-dimensional structure of mostly phospholipids with embedded proteins. This two-dimensional structure plays many key roles in the life of a cell. Transmembrane proteins, for example, play the role of a gate for different ions (such as Ca2+). Also found are peripheral proteins that are used as enzymes for different purposes in the inner leaflet of the plasma membrane. Phospholipids, in particular play three key roles. Firstly, some members of this group are used to store energy. Secondly, the hydrophobic and hydrophilic properties inherent to phospholipids enable them to be used as building blocks of the cell membrane by forming an asymmetric bilayer. This provides a shielding protection against the outer environment while at the same time keeping the organelles and cytosol from leaking out of the cell. Finally lipids are involved in regulating the aggregation of proteins in the membrane. In addition, some subspecies such as phosphatidylinositol (PtdIns) are second messenger molecules in their own right, thus playing an important role in cellular signaling events. In my work presented in this thesis, I am focusing on the role of some phospholipids as signaling molecules and in particular the physicochemical underpinnings that could be used in their spatiotemporal organization in the cellular plasma membrane. I am specifically concerned with the important family of phosphatidylinositol lipids. PtdIns are very well known for their role as signaling molecules in numerous cell events. They are located in the inner leaflet of the plasma membrane as well as part of the membrane of other organelles. Studies of these signaling molecules in their in vivo environment present many challenges: Firstly, the complexity of interactions due to the numerous entities present in eukaryotic cell membranes makes it difficult to establish clear cause and effect relationships. Secondly, due to their size, our inability to probe these

Solid-state NMR paramagnetic relaxation enhancement immersion depth studies in phospholipid bilayers

KAUST Repository

Chu, Shidong

2010-11-01

A new approach for determining the membrane immersion depth of a spin-labeled probe has been developed using paramagnetic relaxation enhancement (PRE) in solid-state NMR spectroscopy. A DOXYL spin label was placed at different sites of 1-palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine (PSPC) phospholipid bilayers as paramagnetic moieties and the resulting enhancements of the longitudinal relaxation (T1) times of 31P nuclei on the surface of the bilayers were measured by a standard inversion recovery pulse sequence. The 31P NMR spin-lattice relaxation times decrease steadily as the DOXYL spin label moves closer to the surface as well as the concentration of the spin-labeled lipids increase. The enhanced relaxation vs. the position and concentration of spin-labels indicate that PRE induced by the DOXYL spin label are significant to determine longer distances over the whole range of the membrane depths. When these data were combined with estimated correlation times τc, the r-6-weighted, time-averaged distances between the spin-labels and the 31P nuclei on the membrane surface were estimated. The application of using this solid-state NMR PRE approach coupled with site-directed spin labeling (SDSL) may be a powerful method for measuring membrane protein immersion depth. © 2010 Elsevier Inc. All rights reserved.

Docosahexaenoic (DHA modulates phospholipid-hydroperoxide glutathione peroxidase (Gpx4 gene expression to ensure self-protection from oxidative damage in hippocampal cells

Directory of Open Access Journals (Sweden)

Veronica eCasañas-Sanchez

2015-07-01

Full Text Available Docosahexaenoic acid (DHA, 22:6n-3 is a unique polyunsaturated fatty acid particularly abundant in nerve cell membrane phospholipids. DHA is a pleiotropic molecule that, not only modulates the physicochemical properties and architecture of neuronal plasma membrane, but it is also involved in multiple facets of neuronal biology, from regulation of synaptic function to neuroprotection and modulation of gene expression. As a highly unsaturated fatty acid due to the presence of six double bonds, DHA is susceptible for oxidation, especially in the highly pro-oxidant environment of brain parenchyma. We have recently reported the ability of DHA to regulate the transcriptional program controlling neuronal antioxidant defenses in a hippocampal cell line, especially the glutathione/glutaredoxin system. Within this antioxidant system, DHA was particularly efficient in triggering the upregulation of Gpx4 gene, which encodes for the nuclear, cytosolic and mitochondrial isoforms of phospholipid-hydroperoxide glutathione peroxidase (PH-GPx/GPx4, the main enzyme protecting cell membranes against lipid peroxidation and capable to reduce oxidized phospholipids in situ. We show here that this novel property of DHA is also significant in the hippocampus of wild-type mice and APP/PS1 transgenic mice, a familial model of Alzheimer’s disease. By doing this, DHA stimulates a mechanism to self-protect from oxidative damage even in the neuronal scenario of high aerobic metabolism and in the presence of elevated levels of transition metals, which inevitably favor the generation of reactive oxygen species. Noticeably, DHA also upregulated a novel Gpx4 splicing variant, harboring part of the first intronic region, which according to the ‘sentinel RNA hypothesis’ would expand the ability of Gpx4 (and DHA to provide neuronal antioxidant defense independently of conventional nuclear splicing in cellular compartments, like dendritic zones, located away from nuclear

Quantification of fatty acids as methyl esters and phospholipids in cheese samples after separation of triacylglycerides and phospholipids

International Nuclear Information System (INIS)

Hauff, Simone; Vetter, Walter

2009-01-01

Determination of the individual fatty acid composition of neutral- and phospholipids as well as the phospholipid content of dairy food and other foodstuffs are important tasks in life sciences. For these purposes, a method was developed for the separation of lipids (standards of triolein and diacylphosphatidylcholines as well as three cheese samples) by solid-phase extraction using a self-packed column filled with partly deactivated silica. Non-halogenated solvents were used for the elution of the lipid classes. Cyclohexane/ethyl acetate (1:1, v/v) served for the elution of neutral lipids, while polar lipids were eluted with three solvents (ethyl acetate/methanol, methanol, and methanol/water) into one fraction. The separated lipid fractions were transesterified and the individual fatty acids were quantified by using gas chromatography coupled to electron ionization mass spectrometry (GC/EI-MS) in the selected ion monitoring (SIM) mode. The recovery rate for standard phosphatidylcholines was ∼90% and cross-contamination from neutral lipids was negligible. The method was applied to cheese samples. Quantitative amounts of individual fatty acids in the phospholipid fraction were eq ) were found to be representative for the average contribution of fatty acids to all classes of phospholipids in dairy products. Using this approach, the phospholipid content of lipids from mozzarella, camembert, and goat cream cheese was 0.60%, 1.42% and 0.79%, respectively

Membrane mimetic surface functionalization of nanoparticles: Methods and applications

Science.gov (United States)

Weingart, Jacob; Vabbilisetty, Pratima; Sun, Xue-Long

2013-01-01

Nanoparticles (NPs), due to their size-dependent physical and chemical properties, have shown remarkable potential for a wide range of applications over the past decades. Particularly, the biological compatibilities and functions of NPs have been extensively studied for expanding their potential in areas of biomedical application such as bioimaging, biosensing, and drug delivery. In doing so, surface functionalization of NPs by introducing synthetic ligands and/or natural biomolecules has become a critical component in regards to the overall performance of the NP system for its intended use. Among known examples of surface functionalization, the construction of an artificial cell membrane structure, based on phospholipids, has proven effective in enhancing biocompatibility and has become a viable alternative to more traditional modifications, such as direct polymer conjugation. Furthermore, certain bioactive molecules can be immobilized onto the surface of phospholipid platforms to generate displays more reminiscent of cellular surface components. Thus, NPs with membrane-mimetic displays have found use in a range of bioimaging, biosensing, and drug delivery applications. This review herein describes recent advances in the preparations and characterization of integrated functional NPs covered by artificial cell membrane structures and their use in various biomedical applications. PMID:23688632

Atomistic Monte Carlo simulation of lipid membranes

DEFF Research Database (Denmark)

Wüstner, Daniel; Sklenar, Heinz

2014-01-01

Biological membranes are complex assemblies of many different molecules of which analysis demands a variety of experimental and computational approaches. In this article, we explain challenges and advantages of atomistic Monte Carlo (MC) simulation of lipid membranes. We provide an introduction...... into the various move sets that are implemented in current MC methods for efficient conformational sampling of lipids and other molecules. In the second part, we demonstrate for a concrete example, how an atomistic local-move set can be implemented for MC simulations of phospholipid monomers and bilayer patches...

The Plasma Membrane of Saccharomyces cerevisiae : Structure, Function, and Biogenesis

NARCIS (Netherlands)

VANDERREST, ME; KAMMINGA, AH; NAKANO, A; ANRAKU, Y; POOLMAN, B; KONINGS, WN

The composition of phospholipids, sphingolipids, and sterols in the plasma membrane has a strong influence on the activity of the proteins associated or embedded in the lipid bilayer. Since most lipid-synthesizing enzymes in Saccharomyces cerevisiae are located in intracellular organelles, an

Sum frequency generation image reconstruction: Aliphatic membrane under spherical cap geometry

Energy Technology Data Exchange (ETDEWEB)

Volkov, Victor [Bereozovaya 2A, Konstantinovo, Moscow Region 140207 (Russian Federation)

2014-10-07

The article explores an opportunity to approach structural properties of phospholipid membranes using Sum Frequency Generation microscopy. To establish the principles of sum frequency generation image reconstruction in such systems, at first approach, we may adopt an idealistic spherical cap uniform assembly of hydrocarbon molecules. Quantum mechanical studies for decanoic acid (used here as a representative molecular system) provide necessary information on transition dipole moments and Raman tensors of the normal modes specific to methyl terminal – a typical moiety in aliphatic (and phospholipid) membranes. Relative degree of localization and frequencies of the normal modes of methyl terminals make nonlinearities of this moiety to be promising in structural analysis using Sum Frequency Generation imaging. Accordingly, the article describes derivations of relevant macroscopic nonlinearities and suggests a mapping procedure to translate amplitudes of the nonlinearities onto microscopy image plane according to geometry of spherical assembly, local molecular orientation, and optical geometry. Reconstructed images indicate a possibility to extract local curvature of bilayer envelopes of spherical character. This may have practical implications for structural extractions in membrane systems of practical relevance.

Storage stability of marine phospholipids emulsions

DEFF Research Database (Denmark)

Lu, Henna Fung Sieng; Nielsen, Nina Skall; Baron, Caroline Pascale

Marine phospholipids (MPL) are believed to provide more advantages than fish oil from the same source. They are considered to have a better bioavailability, a better resistance towards oxidation and a higher content of polyunsaturated fatty acids such as eicosapentaenoic (EPA) and docosahexaenoic...... acids (DHA) than oily triglycerides (fish oil). Therefore, the objective of this study is to explore the feasibility of using marine phospholipids emulsions as delivery system through investigation of the physical, oxidative and hydrolytic stability of MPL emulsions with or without addition of fish oil....... The effect of initial Peroxide Value, total lipids, phospholipids and antioxidants content on stability of MPL emulsions were studied. The physical stability was investigated through measurement of particle size distribution and creaming stability, which involve measurement of changes (%) in emulsion volume...

Fatty Acids in Membranes as Homeostatic, Metabolic and Nutritional Biomarkers: Recent Advancements in Analytics and Diagnostics

Directory of Open Access Journals (Sweden)

Carla Ferreri

2016-12-01

Full Text Available Fatty acids, as structural components of membranes and inflammation/anti-inflammatory mediators, have well-known protective and regulatory effects. They are studied as biomarkers of pathological conditions, as well as saturated and unsaturated hydrophobic moieties in membrane phospholipids that contribute to homeostasis and physiological functions. Lifestyle, nutrition, metabolism and stress—with an excess of radical and oxidative processes—cause fatty acid changes that are examined in the human body using blood lipids. Fatty acid-based membrane lipidomics represents a powerful diagnostic tool for assessing the quantity and quality of fatty acid constituents and also for the follow-up of the membrane fatty acid remodeling that is associated with different physiological and pathological conditions. This review focuses on fatty acid biomarkers with two examples of recent lipidomic research and health applications: (i monounsaturated fatty acids and the analytical challenge offered by hexadecenoic fatty acids (C16:1; and (ii the cohort of 10 fatty acids in phospholipids of red blood cell membranes and its connections to metabolic and nutritional status in healthy and diseased subjects.

Targeted metabolomic profiling indicates structure-based perturbations in serum phospholipids in children with acetaminophen overdose

Directory of Open Access Journals (Sweden)

Sudeepa Bhattacharyya

Full Text Available Phospholipids are an important class of lipids that act as building blocks of biological cell membranes and participate in a variety of vital cellular functions including cell signaling. Previous studies have reported alterations in phosphatidylcholine (PC and lysophosphatidylcholine (lysoPC metabolism in acetaminophen (APAP-treated animals or cell cultures. However, little is known about phospholipid perturbations in humans with APAP toxicity. In the current study, targeted metabolomic analysis of 180 different metabolites including 14 lysoPCs and 73 PCs was performed in serum samples from children and adolescents hospitalized for APAP overdose. Metabolite profiles in the overdose group were compared to those of healthy controls and hospitalized children receiving low dose APAP for treatment of pain or fever (therapeutic group. PCs and lysoPCs with very long chain fatty acids (VLCFAs were significantly decreased in the overdose group, while those with comparatively shorter chain lengths were increased in the overdose group compared to the therapeutic and control groups. All ether linked PCs were decreased in the overdose group compared to the controls. LysoPC-C26:1 was highly reduced in the overdose group and could discriminate between the overdose and control groups with 100% sensitivity and specificity. The PCs and lysoPCs with VLCFAs showed significant associations with changes in clinical indicators of drug metabolism (APAP protein adducts and liver injury (alanine aminotransferase, or ALT. Thus, a structure-dependent reduction in PCs and lysoPCs was observed in the APAP-overdose group, which may suggest a structure-activity relationship in inhibition of enzymes involved in phospholipid metabolism in APAP toxicity. Keywords: Metabolomics, Phospholipids, Acetaminophen, Hepatotoxicity, Drug

Improved stability of highly fluorinated phospholipid-based vesicles in the presence of bile salts.

Science.gov (United States)

Gadras, C; Santaella, C; Vierling, P

1999-01-04

The stability of fluorinated phospholipid-based vesicles in terms of detergent-induced release of encapsulated carboxyfluorescein has been evaluated. The fluorinated liposomes are substantially more resistant towards the lytic action of sodium taurocholate than conventional DSPC or even DSPC/CH 1/1 liposomes. Concerning structure/permeability relationships, the larger the fluorination degree of the membrane, the higher the resistance of the fluorinated liposomes to their destruction by the detergent. These results show that fluorinated liposomes have a promising potential as drug carrier and delivery systems for oral administration.

Enzymatic modification of phospholipids forfunctional applications and human nutrition

DEFF Research Database (Denmark)

Guo, Zheng; Vikbjerg, Anders / Falk; Xu, Xuebing

2005-01-01

analogs based on the latest understanding of pivotal role of phospholipids in manifold biological processes, exploration of remarkable application potentials of phospholipids in meliorating human health, as well as development of new chemical and biotechnological approaches applied to the modification...... design. This will of course provide fundamental bases also for the development of enzymatic technology to produce structured or modified phospholipids....

Novel Antimicrobial Peptide Dendrimers with Amphiphilic Surface and Their Interactions with Phospholipids — Insights from Mass Spectrometry

Directory of Open Access Journals (Sweden)

Paulina Zielinska

2013-06-01

Full Text Available A series of new peptide dendrimers with amphiphilic surface, designed around a dendronized ornithine (Orn core were synthesized and characterized by ESI-MS, 1H-, 13C- NMR, and CD spectrometry. An improved antimicrobial potency against S. aureus and E. coli was detected as a result of an increased charge, higher branching and variable lipophilicity of the residues located at the C-terminus. Minimal inhibitory concentration (MIC values indicated that the selected dendrimers were not sensitive to the physiological concentration of Na+ and K+ ions (100 mM, but expressed reduced potency at 10 mM concentration of Mg2+ and Ca2+ ions. Circular dichroism (CD curves measured under various conditions revealed structure and solvent-dependent curve evolution. ESI-MS studies of gas-phase interactions between selected dendrimers and both anionic (DMPG and neutral (DMPC phospholipids revealed the presence of variously charged dendrimer/phospholipid aggregates with 1:1 to 1:5 stoichiometry. The collision-induced fragmentation (CID of the most abundant [dendrimer/phospholipid]2+ ions of the 1:1 stoichiometry demonstrated that the studied dendrimers formed stronger complexes with anionic DMPG. Both phospholipids have higher affinity towards dendrimers with a more compact structure. Higher differences in CID energy necessary for dissociation of 50% of the complex formed by dendrimers with DMPG vs. DMPC (DCID50 correlate with a lower hemotoxicity. Mass spectrometry results suggest that for a particular group of compounds the DCID50 might be one of the important factors explaining selectivity of antimicrobial peptides and their branched analogs targeting the bacterial membrane. Both circular dichroism and mass spectrometry studies demonstrated that dendrimers of Nα- and Nε-series possess a different conformation in solution and different affinity to model phospholipids, what might influence their specific microbicidal mechanism.

Phospholipid fatty acid and phospholipid etherlipid fingerprints approach to describe complex soil microbial community under impact of cattle husbandry

Czech Academy of Sciences Publication Activity Database

Elhottová, Dana; Němcová, Anna; Gattinger, A.

2007-01-01

Roč. 48, - (2007), s. 73 ISSN 0009-0646. [Kongres Československé společnosti mikrobiologické /24./. 02.10.2007-05.10.2007, Liberec] Institutional research plan: CEZ:AV0Z60660521 Keywords : phospholipid fatty acid * phospholipid etherlipid fingerprints * cattle husbandry Subject RIV: EH - Ecology, Behaviour

Measurement of total phospholipids in urine of patients treated with gentamicin.

Science.gov (United States)

Saunders, D A; Begg, E J; Kirkpatrick, C M; Yeo, J; Graham, G G; Bailey, R R

1997-04-01

The excretion of phospholipids in urine may be a marker of the early renal toxicity of the aminoglycoside antibiotics. Urinary phospholipids are formed in myeloid bodies which develop in the lysosomes of proximal tubules during treatment with the aminoglycosides, and overflow into the urine. Published assays were modified in order to measure the total phospholipid concentrations in human urine. Phospholipids were extracted from freeze-dried urine samples, digested in concentrated sulphuric acid, and the inorganic phosphorus content determined by complexing with ammonium molybdate and measuring the absorbance at 820 nm. Ten septicaemic patients treated with gentamicin for 5-7 days had significantly higher urine phospholipid concentrations than 10 healthy untreated control subjects (P < 0.0001). There was a negative linear relationship between phospholipid excretion and creatinine clearance (r2 = 0.71). In 34 patients with acute pyelonephritis, increased phospholipid concentrations were observed prior to treatment compared with healthy controls (P < 0.001) and did not alter during treatment with gentamicin. However, the phospholipid concentrations decreased significantly after treatment was completed (P < 0.03). These studies suggest that urinary phospholipids may indicate early aminoglycoside toxicity but with poor specificity, as many of the infections being treated may themselves be associated with phospholipiduria.

Rheological properties of poly(vinyl alcohol) (PVA) derived composite membranes for fuel cells

International Nuclear Information System (INIS)

Remiš, T

2017-01-01

Rheological properties of new anhydrous proton conducting membrane based on PVA, tetraethyl orthosilicate (TEOS),sulfosuccinic acid (SSA), titanium dioxide (TiO 2 )was examined at various stoichiometric ratios. SSA was used as sulfonating agents to form a crosslinked structure and as proton source, whereas TEO Sand TiO 2 were utilized to improve the thermal and mechanical properties of the membrane. In order to verify that all the substances were immobilized into the matrix, the membranes were analysed by means of FT-IR. The rheological, mechanical and thermal properties of the membranes were investigated using rheometer ARES G2 and thermogravimetic analyser (TGA).The analysis of mixed PVA solutions exhibited a unique behaviour of viscosity with increased crosslink density. The dynamic storage modulus G´ of dried composite membranes shows better mechanical resistance and increased tolerance to pressure applied during membrane electrode assembly (MEA). (paper)

[Modification of retinal photoreceptor membranes and Ca ion binding].

Science.gov (United States)

Korchagin, V P; Berman, A L; Shukoliukov, S A; Rychkova, M P; Etingof, R N

1978-10-01

Calcium binding by modified photoreceptor membranes of cattle retina has been studied. Ca2+-binding the membranes significantly changes after C-phospholipase treatment, displaying the initial growth (less than 65% of lipid phosphorus removed) with subsequent decrease (more than 65% of phosphorus removed). Liposomes of the photoreceptor membranes lipids were found to bind more calcium than do the native photoreceptor membranes. Proteolytic enzymes (papaine, pronase) splitting some rhodopsin fragments do not affect the ability of the membrane to bind Ca2+. The increase of light-induced Ca-binding is observed only after the outer segments preincubation under conditions providing for rhodopsin phosphorylation. This effect was observed also after the splitting of the rhodopsin fragment by papaine. It is concluded that calcium binding in the photoreceptor membranes is mainly due to the phosphate groups of phospholipids.

Emulsifying triglycerides with dairy phospholipids instead of soy lecithin modulates gut lipase activity

DEFF Research Database (Denmark)

Mathiassen, Jakob Hovalt; Nejrup, Rikke Guldhammer; Frøkiær, Hanne

2015-01-01

in particular to limit fatty acid absorption in babies given infant formulas. Since interaction between the lipid droplet and the gastric and duodenal lipases occur through the hydrophobic/hydrophilic interface, the composition of the emulsifier may be crucial for efficient hydrolysis. We therefore determined...... hydrolytic rate of gastric lipase and pancreatic lipase, on their own or pancreatic lipase after gastric lipase on TAG droplets of similar size emulsified in either soy lecithin (SL) or in bovine milk phospholipids (MPL), more similar to human milk globule membrane lipids than soy lecithin. Gastric lipase...... for formulas for term-born infants....

Association of canalicular membrane enzymes with bile acid micelles and lipid aggregates in human and rat bile.

Science.gov (United States)

Accatino, L; Pizarro, M; Solís, N; Koenig, C S

1995-01-18

This study was undertaken to gain insights into the characteristics of the polymolecular association between canalicular membrane enzymes, bile acids, cholesterol and phospholipids in bile and into the celular mechanisms whereby the enzymes are secreted into bile. With this purpose, we studied the distribution of bile acids, cholesterol, phospholipids, proteins and representative canalicular membrane enzymes (alkaline phosphatase, 5'-nucleotidase and gamma-glutamyl transpeptidase), which can be considered specific marker constituents, in bile fractions enriched in phospholipid-cholesterol lamellar structures (multilamellar and unilamellar vesicles) and bile acid-mixed micelles. These fractions were isolated by ultracentrifugation from human hepatic bile, normal rat bile and bile of rats treated with diosgenin, a steroid that induces a marked increase in biliary cholesterol secretion, and were characterized by density, lipid composition and transmission electron microscopy. These studies demonstrate that alkaline phosphatase, 5'-nucleotidase and gamma-glutamyl transpeptidase are secreted into both human and rat bile where they are preferentially associated with bile acid-mixed micelles, suggesting a role for bile acids in both release of these enzymes and lipids from the canalicular membrane and solubilization in bile. In addition, heterogeneous association of these enzymes with nonmicellar, lamellar structures in human and rat bile is consistent with the hypothesis that processes independent of the detergent effects of bile acids might also result in the release of specific intrinsic membrane proteins into bile.

Key factors regulating the mass delivery of macromolecules to model cell membranes

DEFF Research Database (Denmark)

Campbell, Richard A.; Watkins, Erik B.; Jagalski, Vivien

2014-01-01

We show that both gravity and electrostatics are key factors regulating interactions between model cell membranes and self-assembled liquid crystalline aggregates of dendrimers and phospholipids. The system is a proxy for the trafficking of reservoirs of therapeutic drugs to cell membranes for slow...... of the aggregates to activate endocytosis pathways on specific cell types is discussed in the context of targeted drug delivery applications....

Phospholipid transfer from vesicles to high density lipoproteins, catalyzed by human plasma phospholipid transfer protein

International Nuclear Information System (INIS)

Sweeny, S.A.

1985-01-01

Human plasma phospholipid transfer protein (PLTP) catalyzes the mass transfer of phosphatidylcholine (PC). Partial purification of PLTP yielded proteins with apparent M/sub r/ = 59,000 and 40,000 by SDS-PAGE. PLTP activity was measured by transfer of [ 14 C]L-α-dipalmitoyl PC from egg-PC vesicles to HDL. Activity was enhanced at low pH (4.5) upon addition of β-mercaptoethanol while Ca +2 and Na + had no effect. E/sub act/ for facilitated PC transfer was 18.2 +/- 2 kcal/mol. The donor specificity of PLTP was examined using vesicles containing egg-PC plus cholesterol or sphingomyelin. The fluidity of the donor membrane (measured by fluorescence polarization of diphenylhexatriene) correlated strongly with a decrease in PLTP activity. Phosphatidic acid did not affect activity. Increase in vesicle size reduced activity. The acceptor specificity of PLTP was examined using chemically modified HDL. PLTP activity increased up to 1.7-fold with an initial increase in negative charge and then decreased upon extensive modification. A mechanism is proposed where PLTP binds to vesicls and enhances the diffusion of PC into the medium where it is adsorbed by HDL

Probing Induced Structural Changes in Biomimetic Bacterial Cell Membrane Interactions with Divalent Cations

Energy Technology Data Exchange (ETDEWEB)

Holt, Allison M [ORNL; Standaert, Robert F [ORNL; Jubb, Aaron M [ORNL; Katsaras, John [ORNL; Johs, Alexander [ORNL

2017-01-01

Biological membranes, formed primarily by the self-assembly of complex mixtures of phospholipids, provide a structured scaffold for compartmentalization and structural processes in living cells. The specific physical properties of phospholipid species present in a given membrane play a key role in mediating these processes. Phosphatidylethanolamine (PE), a zwitterionic lipid present in bacterial, yeast, and mammalian cell membranes, is exceptional. In addition to undergoing the standard lipid polymorphic transition between the gel and liquid-crystalline phase, it can also assume an unusual polymorphic state, the inverse hexagonal phase (HII). Divalent cations are among the factors that drive the formation of the HII phase, wherein the lipid molecules form stacked tubular structures by burying the hydrophilic head groups and exposing the hydrophobic tails to the bulk solvent. Most biological membranes contain a lipid species capable of forming the HII state suggesting that such lipid polymorphic structural states play an important role in structural biological processes such as membrane fusion. In this study, the interactions between Mg2+ and biomimetic bacterial cell membranes composed of PE and phosphatidylglycerol (PG) were probed using differential scanning calorimetry (DSC), small-angle x-ray scattering (SAXS), and fluorescence spectroscopy. The lipid phase transitions were examined at varying ratios of PE to PG and upon exposure to physiologically relevant concentrations of Mg2+. An understanding of these basic interactions enhances our understanding of membrane dynamics and how membrane-mediated structural changes may occur in vivo.

Reorganization of Azospirillum brasilense cell membrane is mediated by lipid composition adjustment to maintain optimal fluidity during water deficit.

Science.gov (United States)

Cesari, A B; Paulucci, N S; Biasutti, M A; Reguera, Y B; Gallarato, L A; Kilmurray, C; Dardanelli, M S

2016-01-01

We study the Azospirillum brasilense tolerance to water deficit and the dynamics of adaptive process at the level of the membrane. Azospirillum brasilense was exposed to polyethylene glycol (PEG) growth and PEG shock. Tolerance, phospholipids and fatty acid (FA) composition and membrane fluidity were determined. Azospirillum brasilense was able to grow in the presence of PEG; however, its viability was reduced. Cells grown with PEG showed membrane fluidity similar to those grown without, the lipid composition was modified, increasing phosphatidylcholine and decreasing phosphatidylethanolamine amounts. The unsaturation FAs degree was reduced. The dynamics of the adaptive response revealed a decrease in fluidity 20 min after the addition of PEG, indicating that the PEG has a fluidizing effect on the hydrophobic region of the cell membrane. Fluidity returned to initial values after 60 min of PEG exposure. Azospirillum brasilense is able to perceive osmotic changes by changing the membrane fluidity. This effect is offset by changes in the composition of membrane phospholipid and FA, contributing to the homeostasis of membrane fluidity under water deficit. This knowledge can be used to develop new Azospirillum brasilense formulations showing an adapted membrane to water deficit. © 2015 The Society for Applied Microbiology.

Mechanisms of membrane binding of small GTPase K-Ras4B farnesylated hypervariable region.

Science.gov (United States)

Jang, Hyunbum; Abraham, Sherwin J; Chavan, Tanmay S; Hitchinson, Ben; Khavrutskii, Lyuba; Tarasova, Nadya I; Nussinov, Ruth; Gaponenko, Vadim

2015-04-10

K-Ras4B belongs to a family of small GTPases that regulates cell growth, differentiation and survival. K-ras is frequently mutated in cancer. K-Ras4B association with the plasma membrane through its farnesylated and positively charged C-terminal hypervariable region (HVR) is critical to its oncogenic function. However, the structural mechanisms of membrane association are not fully understood. Here, using confocal microscopy, surface plasmon resonance, and molecular dynamics simulations, we observed that K-Ras4B can be distributed in rigid and loosely packed membrane domains. Its membrane binding domain interaction with phospholipids is driven by membrane fluidity. The farnesyl group spontaneously inserts into the disordered lipid microdomains, whereas the rigid microdomains restrict the farnesyl group penetration. We speculate that the resulting farnesyl protrusion toward the cell interior allows oligomerization of the K-Ras4B membrane binding domain in rigid microdomains. Unlike other Ras isoforms, K-Ras4B HVR contains a single farnesyl modification and positively charged polylysine sequence. The high positive charge not only modulates specific HVR binding to anionic phospholipids but farnesyl membrane orientation. Phosphorylation of Ser-181 prohibits spontaneous farnesyl membrane insertion. The mechanism illuminates the roles of HVR modifications in K-Ras4B targeting microdomains of the plasma membrane and suggests an additional function for HVR in regulation of Ras signaling. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Hydrogen-1 NMR relaxation time studies in membrane: anesthetic systems

International Nuclear Information System (INIS)

Pinto, L.M.A.; Fraceto, L.; Paula, E. de; Franzoni, L.; Spisni, A.

1997-01-01

The study of local anesthetics'(LA) interaction with model phospholipid membranes is justified by the direct correlation between anesthetic's hydrophobicity and its potency/toxicity. By the same reason, uncharged LA species seems to play a crucial role in anesthesia. Most clinically used LA are small amphiphilics with a protonated amine group (pKa around 8). Although both charged (protonated) and uncharged forms can coexist at physiological pH, it has been shown (Lee, Biochim. Biophys. Acta 514:95, 1978; Screier et al. Biochim. Biophys. Acta 769:231, 1984) that the real anesthetic pka can be down-shifted, due to differential partition into membranes, increasing the ratio of uncharged species at pH 7.4. We have measured 1 H-NMR longitudinal relaxation times (T 1 ) for phospholipid and three local anesthetics (tetracaine, lidocaine, benzocaine), in sonicated vesicles at a 3:1 molar ratio. All the LA protons have shown smaller T 1 in this system than in isotropic phases, reflecting LA immobilization caused by insertion in the membrane. T 1 values for the lipid protons in the presence of LA were analyzed, in an attempt to identify specific LA:lipid contact regions. (author)

Composition, structure and properties of POPC–triolein mixtures. Evidence of triglyceride domains in phospholipid bilayers

DEFF Research Database (Denmark)

Duelund, Lars; Jensen, Grethe Vestergaard; Hannibal-Bach, Hans Kristian

2013-01-01

We have in this study investigated the composition, structure and spectroscopical properties of multilamellar vesicles composed of a phospholipid, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), and up to 10mol% of triolein (TO), a triglyceride. We found in agreement with previous results...... as vesicular structures containing entrapped water. Bilayer structure of the membranes was supported by small angle X-ray scattering that showed the membranes to form a lamellar phase. Fluorescence spectroscopy with the polarity sensitive dye Nile red revealed, that the LF samples with more than 5mol......% TO contained pure TO domains. These observations are consistent with an earlier MD simulation study by us and our co-workers suggesting triglycerides to be located in lens shaped, blister-like domains between the two lipid bilayer leaflets (Khandelia et al. (2010) [26])....

78 FR 78393 - Standard on the Storage and Handling of Anhydrous Ammonia; Extension of the Office of Management...

Science.gov (United States)

2013-12-26

...OSHA solicits public comments concerning its proposal to extend OMB approval of the information collection requirements specified in the Storage and Handling of Anhydrous Ammonia Standard (29 CFR 1910.111). Paragraphs (b)(3) and (b)(4) of the Standard have paperwork requirements that apply to non-refrigerated containers and systems and refrigerated containers, respectively; employers use these containers and systems to store and transfer anhydrous ammonia in the workplace.

Using confocal laser scanning microscopy to probe the milk fat globule membrane and associated proteins.

Science.gov (United States)

Gallier, Sophie; Gragson, Derek; Jiménez-Flores, Rafael; Everett, David

2010-04-14

The bovine milk fat globule membrane (MFGM) is an important, biologically relevant membrane due to its functional and health properties. Its composition has been thoroughly studied, but its structure, especially the lateral organization of its components, still remains unclear. We have used confocal laser scanning microscopy (CLSM) to investigate the surface structure of the MFGM in globules with different degrees of processing using two types of fluorescently labeled phospholipid probes and a protein dye. Using this technique, we have observed heterogeneities in the distribution of MFGM lipids and proteins relating to the processing and size of the globules. The effect of pretreating the milk (centrifugation, pasteurization-homogenization and churning) was studied by double-staining the surface of the milk fat globules, followed by observation using CLSM, and by determining the phospholipid profile of raw milk, raw cream, processed milk and buttermilk powder. Our findings agree with other techniques by showing that the composition of the MFGM changes with processing through the loss of phospholipids and the adsorption of caseins and whey proteins onto the surface.

Electroenzymatic Reactions With Oxygen on Laccase-Modified Electrodes in Anhydrous (Pure) Organic Solvent

DEFF Research Database (Denmark)

Yarapolov, A.; Shleev, S.; Zaitseva, E.

2007-01-01

in two different ways: (i) by studying the electroreduction of oxygen in anhydrous DMSO via a direct electron transfer mechanism without proton donors and (ii) by doing the same experiments in the presence of laccase substrates, which display in pure organic solvents both the properties of electron......The electroenzymatic reactions of Trametes hirsuta laccase in the pure organic solvent dimethyl sulfoxide (DMSO) have been investigated within the framework for potential use as a catalytic reaction scheme for oxygen reduction. The bioelectrochemical characteristics of laccase were investigated...... donors as well as the properties of weak acids. The results obtained with laccase in anhydrous DMSO were compared with those obtained previously in aqueous buffer. It was shown that in the absence of proton donors under oxygenated conditions, formation of superoxide anion radicals is prevented at bare...

Impact of Lipid Oxidization on Vertical Structures and Electrostatics of Phospholipid Monolayers Revealed by Combination of Specular X-ray Reflectivity and Grazing-Incidence X-ray Fluorescence.

Science.gov (United States)

Korytowski, Agatha; Abuillan, Wasim; Makky, Ali; Konovalov, Oleg; Tanaka, Motomu

2015-07-30

The influence of phospholipid oxidization of floating monolayers on the structure perpendicular to the global plane and on the density profiles of ions near the lipid monolayer has been investigated by a combination of grazing incidence X-ray fluorescence (GIXF) and specular X-ray reflectivity (XRR). Systematic variation of the composition of the floating monolayers unravels changes in the thickness, roughness and electron density of the lipid monolayers as a function of molar fraction of oxidized phospholipids. Simultaneous GIXF measurements enable one to qualitatively determine the element-specific density profiles of monovalent (K(+) or Cs(+)) and divalent ions (Ca(2+)) in the vicinity of the interface in the presence and absence of two types of oxidized phospholipids (PazePC and PoxnoPC) with high spatial accuracy (±5 Å). We found the condensation of Ca(2+) near carboxylated PazePC was more pronounced compared to PoxnoPC with an aldehyde group. In contrast, the condensation of monovalent ions could hardly be detected even for pure oxidized phospholipid monolayers. Moreover, pure phospholipid monolayers exhibited almost no ion specific condensation near the interface. The quantitative studies with well-defined floating monolayers revealed how the elevation of lipid oxidization level alters the structures and functions of cell membranes.

Large-scale preparation of clove essential oil and eugenol-loaded liposomes using a membrane contactor and a pilot plant.

Science.gov (United States)

Sebaaly, Carine; Greige-Gerges, Hélène; Agusti, Géraldine; Fessi, Hatem; Charcosset, Catherine

2016-01-01

Based on our previous study where optimal conditions were defined to encapsulate clove essential oil (CEO) into liposomes at laboratory scale, we scaled-up the preparation of CEO and eugenol (Eug)-loaded liposomes using a membrane contactor (600 mL) and a pilot plant (3 L) based on the principle of ethanol injection method, both equipped with a Shirasu Porous Glass membrane for injection of the organic phase into the aqueous phase. Homogenous, stable, nanometric-sized and multilamellar liposomes with high phospholipid, Eug loading rates and encapsulation efficiency of CEO components were obtained. Saturation of phospholipids and drug concentration in the organic phase may control the liposome stability. Liposomes loaded with other hydrophobic volatile compounds could be prepared at large scale using the ethanol injection method and a membrane for injection.

The influence of erythrocyte maturity on ion transport and membrane lipid composition in the rat

Czech Academy of Sciences Publication Activity Database

Vokurková, Martina; Rauchová, Hana; Dobešová, Zdenka; Loukotová, Jana; Nováková, O.; Kuneš, Jaroslav; Zicha, Josef

2016-01-01

Roč. 65, č. 1 (2016), s. 91-99 ISSN 0862-8408 R&D Projects: GA MZd(CZ) NV15-25396A; GA ČR(CZ) GAP304/12/0259 Institutional support: RVO:67985823 Keywords : reticulocytes * immature erythrocytes * mean cellular hemoglobin content * membrane phospholipids * membrane cholesterol Subject RIV: ED - Physiology Impact factor: 1.461, year: 2016

Characterization of methanotrophic bacteria on the basis of intact phospholipid profiles.

Science.gov (United States)

Fang, J; Barcelona, M J; Semrau, J D

2000-08-01

The intact phospholipid profiles (IPPs) of seven species of methanotrophs from all three physiological groups, type I, II and X, were determined using liquid chromatography/electrospray ionization/mass spectrometry. In these methanotrophs, two major classes of phospholipids were found, phosphatidylglycerol (PG) and phosphatidylethanolamine (PE) as well as its derivatives phosphatidylmethylethanolamine (PME) and phosphatidyldimethylethanolamine (PDME). Specifically, the type I methanotrophs, Methylomonas methanica, Methylomonas rubra and Methylomicrobium album BG8 were characterized by PE and PG phospholipids with predominantly C16:1 fatty acids. The type II methanotrophs, Methylosinus trichosporium OB3b and CSC1 were characterized by phospholipids of PG, PME and PDME with predominantly C18:1 fatty acids. Methylococcus capsulatus Bath, a representative of type X methanotrophs, contained mostly PE (89% of the total phospholipids). Finally, the IPPs of a recently isolated acidophilic methanotroph, Methylocella palustris, showed it had a preponderance of PME phospholipids with 18:1 fatty acids (94% of total). Principal component analysis showed these methanotrophs could be clearly distinguished based on phospholipid profiles. Results from this study suggest that IPP can be very useful in bacterial chemotaxonomy.

Role of phospholipids in the pathophysiology of the gut-liver axis

NARCIS (Netherlands)

Petruzzelli, M.

2010-01-01

Phospholipids represent essential components of bile. Together with bile acids and cholesterol, phospholipids form “mixed micelles”. If sufficient amounts of phospholipids are available, no simple bile acid micelles are present, with prevention of bile acid toxicity and cholesterol crystallization.

Phospholipids of New Zealand Edible Brown Algae.

Science.gov (United States)

Vyssotski, Mikhail; Lagutin, Kirill; MacKenzie, Andrew; Mitchell, Kevin; Scott, Dawn

2017-07-01

Edible brown algae have attracted interest as a source of beneficial allenic carotenoid fucoxanthin, and glyco- and phospholipids enriched in polyunsaturated fatty acids. Unlike green algae, brown algae contain no or little phosphatidylserine, possessing an unusual aminophospholipid, phosphatidyl-O-[N-(2-hydroxyethyl) glycine], PHEG, instead. When our routinely used technique of 31 P-NMR analysis of phospholipids was applied to the samples of edible New Zealand brown algae, a number of signals corresponding to unidentified phosphorus-containing compounds were observed in total lipids. NI (negative ion) ESI QToF MS spectra confirmed the presence of more familiar phospholipids, and also suggested the presence of PHEG or its isomers. The structure of PHEG was confirmed by comparison with a synthetic standard. An unusual MS fragmentation pattern that was also observed prompted us to synthesise a number of possible candidates, and was found to follow that of phosphatidylhydroxyethyl methylcarbamate, likely an extraction artefact. An unexpected outcome was the finding of ceramidephosphoinositol that has not been reported previously as occurring in brown algae. An uncommon arsenic-containing phospholipid has also been observed and quantified, and its TLC behaviour studied, along with that of the newly synthesised lipids.

Effect of acetone accumulation on structure and dynamics of lipid membranes studied by molecular dynamics simulations.

Science.gov (United States)

Posokhov, Yevgen O; Kyrychenko, Alexander

2013-10-01

The modulation of the properties and function of cell membranes by small volatile substances is important for many biomedical applications. Despite available experimental results, molecular mechanisms of action of inhalants and organic solvents, such as acetone, on lipid membranes remain not well understood. To gain a better understanding of how acetone interacts with membranes, we have performed a series of molecular dynamics (MD) simulations of a POPC bilayer in aqueous solution in the presence of acetone, whose concentration was varied from 2.8 to 11.2 mol%. The MD simulations of passive distribution of acetone between a bulk water phase and a lipid bilayer show that acetone favors partitioning into the water-free region of the bilayer, located near the carbonyl groups of the phospholipids and at the beginning of the hydrocarbon core of the lipid membrane. Using MD umbrella sampling, we found that the permeability barrier of ~0.5 kcal/mol exists for acetone partitioning into the membrane. In addition, a Gibbs free energy profile of the acetone penetration across a bilayer demonstrates a favorable potential energy well of -3.6 kcal/mol, located at 15-16Å from the bilayer center. The analysis of the structural and dynamics properties of the model membrane revealed that the POPC bilayer can tolerate the presence of acetone in the concentration range of 2.8-5.6 mol%. The accumulation of the higher acetone concentration of 11.2 mol% results, however, in drastic disordering of phospholipid packing and the increase in the membrane fluidity. The acetone molecules push the lipid heads apart and, hence, act as spacers in the headgroup region. This effect leads to the increase in the average headgroup area per molecule. In addition, the acyl tail region of the membrane also becomes less dense. We suggest, therefore, that the molecular mechanism of acetone action on the phospholipid bilayer has many common features with the effects of short chain alcohols, DMSO, and

A New Thickener for CO2 Anhydrous Fracturing Fluid

Directory of Open Access Journals (Sweden)

Zhang Jian

2015-01-01

Full Text Available CO2 dry fracturing technology is well-known for its advantages. Little water is used in this technology, which is able to ease the pressure of consumption on water resources. Many abroad theoretical researches, laboratory experiments and field tests have been taken to explore the yield mechanism, the adaptability and the technology of pure liquid CO2 fracturing. These achievements have been applied to a variety of reservoirs transformation and improven the effectiveness of stimulation treatment in a degree. The researches and studies in the domestic didn’t get popular until recent years. Thus, this article firstly introduces the main development and application about pure CO2 anhydrous fracturing technology, and sums up the effect and evaluation of its fluid through application examples both in the domestic and abroad. However, although this technology has many excellent qualities, but systematic studies indicate that its proppant-carrying capacity is less competitive because of the low viscosity of pure CO2 liquid and other reasons. In a consequence, it is necessary to develop an appropriate thickener for CO2 anhydrous fracturing fluid to improve its carrying capacity. Then this article describes some studies of previous scholars about CO2 thickener. Then we put forward our own research ideas and transform it into actual experiments. Thanks to the valid performances of these tests, we successfully develop a thickener X and cosolvent B.

Interactions between the intermediate filaments of vimentin and natural or artificial lipid membranes

International Nuclear Information System (INIS)

Perides, G.

1986-01-01

The study has provided evidence to prove that intermediate filaments (IF) are not only encountered in the vicinity of several cellular membrane systems but even become attached to those membranes by stable mechanical bonds. Studies using photoaffinity markers permitted to show in vivo that vimentin occurs in the immediate neighbourhood of membrane lipids. Titration of cellular membranes with radioactively labelled vimentin and desmin pointed to the fact that there is a large excess of acceptor molecules for IF proteins, from which it was concluded that vimentin directly binds to the lipids. This is also consistent with the finding that vesicles made up of cellular lipids readily bind to vimentin filaments and may even interfere with the formation of the latter. The highest vimentin affinity was observed for negatively charged phospholipids, which led to the theory that the association of IF and cellular membranes is firstly attributable to an interaction between the positive N-terminals of IF proteins and upper polar groups of negative phospholipids. The binding of vimentin to cellular mebranes changes under the influence of cellular growth processes and extracellular factors. This was also suggested by the reduced amounts of membrane-bound vimentin found after the incubation of cells in a serum-free medium and the prompt increases in the vimentin content of those membranes, after serum was added. This is one example, among several others, to show that the reactions between IF and cellular membranes are of a reversible nature and controlled and shaped by the cell itself. (orig./MG) [de

Thermal decomposition of anhydrous zinc and cadmium salicylates

International Nuclear Information System (INIS)

Kharitonov, Yu.Ya.; Tujebakhova, Z.K.

1984-01-01

On the basis of studying thermograms, thermogravigrams, IR absorption spectra, X-rayograms of anhydrous znc and cadmium salicylate complexes of the M(HSal) 2 composition, (where M=Zn, Cd; HSal is an anion of once deprotonated salicyclic acid H 2 Sal) and products of their thermal transformations, the processes are characterized of stage-by-stage thermal decomposition of these compounds under continuous heating in the air from room temperature to approximately 1000 deg C. It is shown that the Cd(HSal) 2 pyrolysis proceeds with the formation of CdSal at 170-250 deg C and CdO - at 320-460 deg C

An Investigation of Proton Conductivity of Vinyltriazole-Grafted PVDF Proton Exchange Membranes Prepared via Photoinduced Grafting

Directory of Open Access Journals (Sweden)

Sinan Sezgin

2014-01-01

Full Text Available Proton exchange membrane fuel cells (PEMFCs are considered to be a promising technology for clean and efficient power generation in the twenty-first century. In this study, high performance of poly(vinylidene fluoride (PVDF and proton conductivity of poly(1-vinyl-1,2,4-triazole (PVTri were combined in a graft copolymer, PVDF-g-PVTri, by the polymerization of 1-vinyl-1,2,4-triazole on a PVDF based matrix under UV light in one step. The polymers were doped with triflic acid (TA at different stoichiometric ratios with respect to triazole units and the anhydrous polymer electrolyte membranes were prepared. All samples were characterized by FTIR and 1H-NMR spectroscopies. Their thermal properties were examined by thermogravimetric analysis (TGA and differential scanning calorimetry (DSC. TGA demonstrated that the PVDF-g-PVTri and PVDF-g-PVTri-(TAx membranes were thermally stable up to 390°C and 330°C, respectively. NMR and energy dispersive X-ray spectroscopy (EDS results demonstrated that PVDF-g-PVTri was successfully synthesized with a degree of grafting of 21%. PVDF-g-PVTri-(TA3 showed a maximum proton conductivity of 6×10-3 Scm−1 at 150°C and anhydrous conditions. CV study illustrated that electrochemical stability domain for PVDF-g-PVTri-(TA3 extended over 4.0 V.

Phospholipase D catalyzes phospholipid metabolism in chemotactic peptide-stimulated HL-60 granulocytes

International Nuclear Information System (INIS)

Pai, J.K.; Siegel, M.I.; Egan, R.W.; Billah, M.M.

1988-01-01

There exists circumstantial evidence for activation of phospholipase D (PLD) in intact cells. However, because of the complexity of phospholipid remodeling processes, it is essential to distinguish PLD clearly from other phospholipases and phospholipid remodeling enzymes. Therefore, to establish unequivocally PLD activity in dimethyl sulfoxide-differentiated HL-60 granulocytes, to demonstrate the relative contribution of PLD to phospholipid turnover, and to validate the hypothesis that the formation of phosphatidylethanol is an expression of PLD-catalyzed transphosphatidylation, we have developed methodologies to label HL-60 granulocytes in 1-O-alkyl-2-acyl-sn-glycero-3-phosphocholine (alkyl-PC) with 32P without labeling cellular ATP. These methodologies involve (a) synthesis of alkyl-lysoPC containing 32P by a combination of enzymatic and chemical procedures and (b) incubation of HL-60 granulocytes with this alkyl-[32P] lysoPC which enters the cell and becomes acylated into membrane-associated alkyl-[32P]PC. Upon stimulation of these 32P-labeled cells with the chemotactic peptide, N-formyl-Met-Leu-Phe (fMLP), alkyl-[32P]phosphatidic acid (alkyl-[32P]PA) is formed rapidly. Because, under these conditions, cellular ATP has not been labeled with 32P, alkyl-[32P]PA must be formed via PLD-catalyzed hydrolysis of alkyl-[32P]PC at the terminal phosphodiester bond. This result conclusively demonstrates fMLP-induced activation of PLD in HL-60 granulocytes. These 32P-labeled HL-60 granulocytes have also been stimulated in the presence of ethanol to produce alkyl-[32P]phosphatidylethanol (alkyl-[32P]PEt). Formation of alkyl-[32P]PEt parallels that of alkyl-[32P]PA with respect to time course, fMLP concentration, inhibition by a specific fMLP antagonist (t-butoxycarbonyl-Met-Leu-Phe), and Ca2+ concentration

Relationship between Al content and substitution mechanism of Al-bearing anhydrous bridgmanites

Science.gov (United States)

Noda, M.; Inoue, T.; Kakizawa, S.

2017-12-01

It is considered that two substitution mechanisms, Tschermak substitution and oxygen vacancy substitution, exist in MgSiO3 bridgmanite for the incorporation of Al in anhydrous condition. Kubo and Akaogi (2000) has conducted the phase equilibrium experiment in the system MgSiO3-Al2O3, and established the phase diagram up to 28 GPa. However the careful observation in the bridgmanite shows that the chemical compositions are slightly deviated from Tschermak substitution join. The same tendency can be also observed in the run products by Irifune et al. (1996). This result indicates that pure Tschermak substitution bridgmanite cannot be stable even in the MgSiO3-Al2O3 join experiment. However, the previous studies used powder samples as the starting materials, so the absorbed water may affect the results. Therefore, we tried to conduct the experiment in the join MgSiO3-Al2O3 in extremely anhydrous condition to clarify whether the pure Tschermak substitution bridgmanite can be stable or not. In addition, we also examined the stability of oxygen vacancy bridgmanite in the extremely anhydrous condition for the comparison. The high pressure synthesis experiments were conducted at 28 GPa and 1600-1700° for 1hour using a Kawai-type multi-anvil apparatus. Four different Al content samples were prepared as the starting materials along the ideal substitution line of Tschermak (Al=0.025, 0.05, 0.1, 0.15 mol) and oxygen-vacancy (Al=0.025, 0.05, 0.075, 0.1 mol) substitutions, respectively (when total cation of 2). The glass rods were used as the starting materials to eliminate the absorbed water on the sample surface. The chemical compositions of the synthesized bridgmanite could not be measured by EPMA because of small grain size less than submicron. Therefore the chemical compositions were estimated from the result of the XRD pattern by subtracting the amount of the other phases. The estimated chemical compositions of Tschermak substitution bridgmanites were consistent with the

Anionic lipids and the maintenance of membrane electrostatics in eukaryotes.

Science.gov (United States)

Platre, Matthieu Pierre; Jaillais, Yvon

2017-02-01

A wide range of signaling processes occurs at the cell surface through the reversible association of proteins from the cytosol to the plasma membrane. Some low abundant lipids are enriched at the membrane of specific compartments and thereby contribute to the identity of cell organelles by acting as biochemical landmarks. Lipids also influence membrane biophysical properties, which emerge as an important feature in specifying cellular territories. Such parameters are crucial for signal transduction and include lipid packing, membrane curvature and electrostatics. In particular, membrane electrostatics specifies the identity of the plasma membrane inner leaflet. Membrane surface charges are carried by anionic phospholipids, however the exact nature of the lipid(s) that powers the plasma membrane electrostatic field varies among eukaryotes and has been hotly debated during the last decade. Herein, we discuss the role of anionic lipids in setting up plasma membrane electrostatics and we compare similarities and differences that were found in different eukaryotic cells.

Radiation-induced lipid peroxidation: influence of oxygen concentration and membrane lipid composition

International Nuclear Information System (INIS)

Wolters, H.; Tilburg, C.A.M. van; Konings, A.W.T.

1987-01-01

Radiation -induced lipid peroxidation phospholipid liposomes was investigated in terms of its dependence on lipid composition and oxygen concentration. Non-peroxidizable lipid incorporated in the liposomes reduced the rate of peroxidation of the peroxidizable phospholipid acyl chains, possibly by restricting the length of chain reactions. The latter effect is believed to be caused by interference of the non-peroxidizable lipids in the bilayer. At low oxygen concentration lipid peroxidation was reduced. The cause of this limited peroxidation may be a reduced number of radical initiation reactions possibly involving oxygen-derived superoxide radicals. Killing of proliferating mammalian cells, irradiated at oxygen concentrations ranging from 0 to 100%, appeared to be independent of the concentration of peroxidizable phospholipids in the cell membranes. This indicates that lipid peroxidation is not the determining process in radiation-induced reproductive cell death. (author)

Mediterranean-style diet effect on the structural properties of the erythrocyte cell membrane of hypertensive patients: the Prevencion con Dieta Mediterranea Study.

Science.gov (United States)

Barceló, Francisca; Perona, Javier S; Prades, Jesús; Funari, Sérgio S; Gomez-Gracia, Enrique; Conde, Manuel; Estruch, Ramon; Ruiz-Gutiérrez, Valentina

2009-11-01

A currently ongoing randomized trial has revealed that the Mediterranean diet, rich in virgin olive oil or nuts, reduces systolic blood pressure in high-risk cardiovascular patients. Here, we present a structural substudy to assess the effect of a Mediterranean-style diet supplemented with nuts or virgin olive oil on erythrocyte membrane properties in 36 hypertensive participants after 1 year of intervention. Erythrocyte membrane lipid composition, structural properties of reconstituted erythrocyte membranes, and serum concentrations of inflammatory markers are reported. After the intervention, the membrane cholesterol content decreased, whereas that of phospholipids increased in all of the dietary groups; the diminishing cholesterol:phospholipid ratio could be associated with an increase in the membrane fluidity. Moreover, reconstituted membranes from the nuts and virgin olive oil groups showed a higher propensity to form a nonlamellar inverted hexagonal phase structure that was related to an increase in phosphatidylethanolamine lipid class. These data suggest that the Mediterranean-style diet affects the lipid metabolism that is altered in hypertensive patients, influencing the structural membrane properties. The erythrocyte membrane modulation described provides insight in the structural bases underlying the beneficial effect of a Mediterranean-style diet in hypertensive subjects.

Ab initio and DFT studies of the structure and vibrational spectra of anhydrous caffeine

Science.gov (United States)

Srivastava, Santosh K.; Singh, Vipin B.

2013-11-01

Vibrational spectra and molecular structure of anhydrous caffeine have been systematically investigated by second order Moller-Plesset (MP2) perturbation theory and density functional theory (DFT) calculations. Vibrational assignments have been made and many previous ambiguous assignments in IR and Raman spectra are amended. The calculated DFT frequencies and intensities at B3LYP/6-311++G(2d,2p) level, were found to be in better agreement with the experimental values. It was found that DFT with B3LYP functional predicts harmonic vibrational wave numbers more close to experimentally observed value when it was performed on MP2 optimized geometry rather than DFT geometry. The calculated TD-DFT vertical excitation electronic energies of the valence excited states of anhydrous caffeine are found to be in consonance to the experimental absorption peaks.

Nanoparticle self-assembly in mixtures of phospholipids with styrene/maleic acid copolymers or fluorinated surfactants

Science.gov (United States)

Vargas, Carolyn; Arenas, Rodrigo Cuevas; Frotscher, Erik; Keller, Sandro

2015-12-01

Self-assembling nanostructures in aqueous mixtures of bilayer-forming lipids and micelle-forming surfactants are relevant to in vitro studies on biological and synthetic membranes and membrane proteins. Considerable efforts are currently underway to replace conventional detergents by milder alternatives such as styrene/maleic acid (SMA) copolymers and fluorinated surfactants. However, these compounds and their nanosized assemblies remain poorly understood as regards their interactions with lipid membranes, particularly, the thermodynamics of membrane partitioning and solubilisation. Using 19F and 31P nuclear magnetic resonance spectroscopy, static and dynamic light scattering, and isothermal titration calorimetry, we have systematically investigated the aggregational state of a zwitterionic bilayer-forming phospholipid upon exposure to an SMA polymer with a styrene/maleic acid ratio of 3 : 1 or to a fluorinated octyl phosphocholine derivative called F6OPC. The lipid interactions of SMA(3 : 1) and F6OPC can be thermodynamically conceptualised within the framework of a three-stage model that treats bilayer vesicles, discoidal or micellar nanostructures, and the aqueous solution as distinct pseudophases. The exceptional solubilising power of SMA(3 : 1) is reflected in very low membrane-saturating and solubilising polymer/lipid molar ratios of 0.10 and 0.15, respectively. Although F6OPC saturates bilayers at an even lower molar ratio of 0.031, this nondetergent does not solubilise lipids even at >1000-fold molar excess, thus highlighting fundamental differences between these two types of mild membrane-mimetic systems. We rationalise these findings in terms of a new classification of surfactants based on bilayer-to-micelle transfer free energies and discuss practical implications for membrane-protein research.Self-assembling nanostructures in aqueous mixtures of bilayer-forming lipids and micelle-forming surfactants are relevant to in vitro studies on biological and

Screening for the drug-phospholipid interaction: correlation to phospholipidosis

DEFF Research Database (Denmark)

Alakoskela, Juha-Matti; Vitovic, Pavol; Kinnunen, Paavo K J

2009-01-01

Phospholipid bilayers represent a complex, anisotropic environment fundamentally different from bulk oil or octanol, for instance. Even "simple" drug association to phospholipid bilayers can only be fully understood if the slab-of-hydrocarbon approach is abandoned and the complex, anisotropic...

Enhanced resolution of membranes in cultured cells by cryoimmobilization and freeze-substitution.

Science.gov (United States)

Wild, P; Schraner, E M; Adler, H; Humbel, B M

2001-05-15

Investigations of cellular processes demand immediate arresting of the process at any given time and excellent retention of cellular material and excellent visibility of membranes. To achieve this goal we used cryofixation to arrest cellular processes instantly and tested diverse freeze-substitution protocols. Madin-Darby kidney cells and Vero cells were grown on carbon-coated sapphire disks. For cryofixation the sapphire disks covered with a cell monolayer were injected with the aid of a guillotine into liquid propane or ethane or a mixture of both cooled by liquid nitrogen. Freezing of the cryogen was prevented by using a partially insulated cylinder and by vigorous stirring that results in a substantial decrement of the freezing point of the cryogen. Cell monolayers can be cryofixed successfully using the guillotine in a safety hood at ambient temperature and humidity or at 37 degrees C and 45% humidity. The freezing unit can also be placed in a laminar flow for working under biohazard conditions. For visualizing cell membranes at high contrast and high resolution, cells were substituted in the presence of various concentrations of glutaraldehyde and osmium tetroxide and the temperature was raised to diverse final temperatures. Substitution for 4 hours at -90 degrees C in anhydrous acetone containing 0.25% anhydrous glutaraldehyde and 0.5% osmium tetroxide followed by a temperature rise of 5 degrees C/hour to 0 degrees C and final incubation for 1 hour at 0 degrees C resulted in high contrast and excellent visibility of subcellular components at the level of the membrane bilayer. The high spatial and temporal resolution makes this methodology an excellent tool for studying cell membrane-bound processes, such as virus-cell interactions. Copyright 2001 Wiley-Liss, Inc.

ROLE OF MEMBRANE LIPID-COMPOSITION IN THE CYTOTOXICITY OF THE SESQUITERPENE LACTONE EUPATORIOPICRIN

NARCIS (Netherlands)

VANDERLINDE, JCC; WOERDENBAG, HJ; MALINGRE, TM; KAMPINGA, HH; KONINGS, AWT

1993-01-01

The aim of the present study was to investigate a possible role of lipid peroxidation in the cytotoxicity of eupatoriopicrin, the principal sesquiterpene lactone from Eupatorum cannabinum L. Incorporation of arachidonic acid acyl chains in the phospholipids of cellular membranes of mouse fibroblast

Structure of the Na,K-ATPase regulatory protein FXYD2b in micelles: implications for membrane-water interfacial arginines.

Science.gov (United States)

Gong, Xiao-Min; Ding, Yi; Yu, Jinghua; Yao, Yong; Marassi, Francesca M

2015-01-01

FXYD2 is a membrane protein responsible for regulating the function of the Na,K-ATPase in mammalian kidney epithelial cells. Here we report the structure of FXYD2b, one of two splice variants of the protein, determined by NMR spectroscopy in detergent micelles. Solid-state NMR characterization of the protein embedded in phospholipid bilayers indicates that several arginine side chains may be involved in hydrogen bond interactions with the phospholipid polar head groups. The structure and the NMR data suggest that FXYD2b could regulate the Na,K-ATPase by modulating the effective membrane surface electrostatics near the ion binding sites of the pump. Copyright © 2014 Elsevier B.V. All rights reserved.

Fabrication Procedures and Birefringence Measurements for Designing Magnetically Responsive Lanthanide Ion Chelating Phospholipid Assemblies.

Science.gov (United States)

Isabettini, Stéphane; Baumgartner, Mirjam E; Fischer, Peter; Windhab, Erich J; Liebi, Marianne; Kuster, Simon

2018-01-03

Bicelles are tunable disk-like polymolecular assemblies formed from a large variety of lipid mixtures. Applications range from membrane protein structural studies by nuclear magnetic resonance (NMR) to nanotechnological developments including the formation of optically active and magnetically switchable gels. Such technologies require high control of the assembly size, magnetic response and thermal resistance. Mixtures of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and its lanthanide ion (Ln 3+ ) chelating phospholipid conjugate, 1,2-dimyristoyl-sn-glycero-3-phospho-ethanolamine-diethylene triaminepentaacetate (DMPE-DTPA), assemble into highly magnetically responsive assemblies such as DMPC/DMPE-DTPA/Ln 3+ (molar ratio 4:1:1) bicelles. Introduction of cholesterol (Chol-OH) and steroid derivatives in the bilayer results in another set of assemblies offering unique physico-chemical properties. For a given lipid composition, the magnetic alignability is proportional to the bicelle size. The complexation of Ln 3+ results in unprecedented magnetic responses in terms of both magnitude and alignment direction. The thermo-reversible collapse of the disk-like structures into vesicles upon heating allows tailoring of the assemblies' dimensions by extrusion through membrane filters with defined pore sizes. The magnetically alignable bicelles are regenerated by cooling to 5 °C, resulting in assembly dimensions defined by the vesicle precursors. Herein, this fabrication procedure is explained and the magnetic alignability of the assemblies is quantified by birefringence measurements under a 5.5 T magnetic field. The birefringence signal, originating from the phospholipid bilayer, further enables monitoring of polymolecular changes occurring in the bilayer. This simple technique is complementary to NMR experiments that are commonly employed to characterize bicelles.

Co-assembly of chitosan and phospholipids into hybrid hydrogels

DEFF Research Database (Denmark)

Mendes, Ana Carina Loureiro; Shekarforoush, Elhamalsadat; Engwer, Christoph

2016-01-01

Novel hybrid hydrogels were formed by adding chitosan (Ch) to phospholipids (P) self-assembled particles in lactic acid. The effect of the phospholipid concentration on the hydrogel properties was investigated and was observed to affect the rate of hydrogel formation and viscoelastic properties...

Effect of Omega-3 Fatty Acids on Erythrocyte Membrane in Diabetic Rats

OpenAIRE

Hussein, Jihan; Mostafa, Ehab; El-Waseef, Maha; El-Khayat, Zakarya; Badawy, Ehsan; Medhat, Dalia

2011-01-01

Background: Diabetes mellitus is a metabolic disease characterized by chronic hyperglycemia resulting from defects in insulin secretion, almost always with a major contribution from insulin resistance which may be affected by cell membrane fatty acids and phospholipids fractions.Aim: To evaluate the effects of omega-3 fatty acids on erythrocyte membrane and also in decreasing oxidative stress in diabetic rats.Material and Methods: Sixty healthy male albino rats weighting 180-200 g divided int...

Modification of trout sperm membranes associated with activation and cryopreservation. Implications for fertilizing potential

Science.gov (United States)

Abstract We investigated the effects of two trout sperm activation solutions on sperm physiology and membrane organization prior to and following cryopreservation using flow cytometry and investigated their impact on in vitro fertility. Cryopreservation caused greater phospholipid disorder (high pl...

The interaction of insulin, glucose, and insulin-glucose mixtures with a phospholipid monolayer.

Science.gov (United States)

Shigenobu, Hayato; McNamee, Cathy E

2012-12-15

We determined how glucose or insulin interacts with a phospholipid monolayer at the air/water interface and explained these mechanisms from a physico-chemical point of view. The 1,2-dipalmitoyl-2-sn-glycero-3-phosphatidylcholine (DPPC) monolayer at an air/water interface acted as a model membrane, which allowed the effect of the molecular packing density in the monolayer on the interactions to be determined. The interaction of glucose, insulin, and a mixture of glucose and insulin to the DPPC monolayer were investigated via surface pressure-area per molecule Langmuir isotherms and fluorescence microscopy. Glucose adsorbed to the underside of the DPPC monolayer, while insulin was able to penetrate through the monolayer when the phospholipid molecules were not densely packed. The presence of a mixture of insulin and glucose affected the molecular packing in the DPPC monolayer differently than the pure insulin or glucose solutions, and the glucose-insulin mixture was seen to be able to penetrate through the monolayer. These results indicated that glucose and insulin interact with one another, giving a material that may then transported through a pore in the monolayer or through the spaces between the molecules of the monolayer. Copyright © 2012 Elsevier Inc. All rights reserved.

Protection of DPPC phospholipid liposomal membrane against radiation oxidative damage by antioxidants

Energy Technology Data Exchange (ETDEWEB)

Marathe, D.L.; Pandey, B.N.; Mishra, K.P [Bhabha Atomic Research Centre, Mumbai (India)

2000-05-01

Investigations in our laboratory on egg lecithin liposomes have recently showed a marked protection against damage by gamma radiation when cholesterol was present in the composition of vesicles suggesting a role of bilayer molecular architecture in the mechanism of free radical mediated lipid peroxidation. Present study was designed to determine the changes in bilayer permeability in DPPC unilamelar vesicles after exposure to gamma radiation by monitoring the leakage of pre-loaded carboxyfluorescein (CF), a marker loaded in aqueous interior of vesicle and fluidity alterations in the bilayer using fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH), a membrane bilayer probe. It was found that radiation doses of an order of magnitude higher were required to produce detectable changes in vesicles of DPPC than in the vesicles of egg lecithin suggesting a modulating role of chemical nature of composition in the membrane radiation sensitivity. It was significant to find that the leakage of CF from and incorporation of DPH into vesicle bilayer showed similar response pattern to radiation doses (0.1-6 kGy) which was also found to be dose rate dependent. Presence of antioxidants; alpha-tocopherol (0.15 mole %) in the bilayer membrane or ascorbic acid (0.1 mM) in the aqueous region significantly protected DPPC vesicles from radiation damage as determined from DPH uptake kinetics suggesting involvement of reactive free radicals of lipids as well as water radicals in the mechanism of membrane peroxidative damage. The magnitude of protection was found to increase with the increasing concentration of both these antioxidants but comparisons showed that {alpha}-tocopherol was far more effective in protecting the vesicles than ascorbic acid. These results contribute to our understanding of the mechanism of radiation oxidative damage and its modification by radical scavenging and/or organizational modulation which emphasize the importance of structure and composition of

Protection of DPPC phospholipid liposomal membrane against radiation oxidative damage by antioxidants

International Nuclear Information System (INIS)

Marathe, D.L.; Pandey, B.N.; Mishra, K.P

2000-01-01

Investigations in our laboratory on egg lecithin liposomes have recently showed a marked protection against damage by gamma radiation when cholesterol was present in the composition of vesicles suggesting a role of bilayer molecular architecture in the mechanism of free radical mediated lipid peroxidation. Present study was designed to determine the changes in bilayer permeability in DPPC unilamelar vesicles after exposure to gamma radiation by monitoring the leakage of pre-loaded carboxyfluorescein (CF), a marker loaded in aqueous interior of vesicle and fluidity alterations in the bilayer using fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH), a membrane bilayer probe. It was found that radiation doses of an order of magnitude higher were required to produce detectable changes in vesicles of DPPC than in the vesicles of egg lecithin suggesting a modulating role of chemical nature of composition in the membrane radiation sensitivity. It was significant to find that the leakage of CF from and incorporation of DPH into vesicle bilayer showed similar response pattern to radiation doses (0.1-6 kGy) which was also found to be dose rate dependent. Presence of antioxidants; alpha-tocopherol (0.15 mole %) in the bilayer membrane or ascorbic acid (0.1 mM) in the aqueous region significantly protected DPPC vesicles from radiation damage as determined from DPH uptake kinetics suggesting involvement of reactive free radicals of lipids as well as water radicals in the mechanism of membrane peroxidative damage. The magnitude of protection was found to increase with the increasing concentration of both these antioxidants but comparisons showed that α-tocopherol was far more effective in protecting the vesicles than ascorbic acid. These results contribute to our understanding of the mechanism of radiation oxidative damage and its modification by radical scavenging and/or organizational modulation which emphasize the importance of structure and composition of

Simulations of simple linoleic acid-containing lipid membranes and models for the soybean plasma membranes.

Science.gov (United States)

Zhuang, Xiaohong; Ou, Anna; Klauda, Jeffery B

2017-06-07

The all-atom CHARMM36 lipid force field (C36FF) has been tested with saturated, monounsaturated, and polyunsaturated lipids; however, it has not been validated against the 18:2 linoleoyl lipids with an unsaturated sn-1 chain. The linoleoyl lipids are common in plants and the main component of the soybean membrane. The lipid composition of soybean plasma membranes has been thoroughly characterized with experimental studies. However, there is comparatively less work done with computational modeling. Our molecular dynamics (MD) simulation results show that the pure linoleoyl lipids, 1-stearoyl-2-linoleoyl-sn-glycero-3-phosphocholine (18:0/18:2) and 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (di-18:2), agree very well with the experiments, which demonstrates the accuracy of the C36FF for the computational study of soybean membranes. Based on the experimental composition, the soybean hypocotyl and root plasma membrane models are developed with each containing seven or eight types of linoleoyl phospholipids and two types of sterols (sitosterol and stigmasterol). MD simulations are performed to characterize soybean membranes, and the hydrogen bonds and clustering results demonstrate that the lipids prefer to interact with the lipids of the same/similar tail unsaturation. All the results suggest that these two soybean membrane models can be used as a basis for further research in soybean and higher plant membranes involving membrane-associated proteins.

Nanomechanics of electrospun phospholipid fiber

Energy Technology Data Exchange (ETDEWEB)

Mendes, Ana C., E-mail: anac@food.dtu.dk, E-mail: ioach@food.dtu.dk; Chronakis, Ioannis S., E-mail: anac@food.dtu.dk, E-mail: ioach@food.dtu.dk [Technical University of Denmark, DTU-Food, Søltofts Plads B227, DK-2800, Kgs. Lyngby (Denmark); Nikogeorgos, Nikolaos; Lee, Seunghwan [Department of Mechanical Engineering, Technical University of Denmark, DK-2800 Kgs. Lyngby (Denmark)

2015-06-01

Electrospun asolectin phospholipid fibers were prepared using isooctane as a solvent and had an average diameter of 6.1 ± 2.7 μm. Their mechanical properties were evaluated by nanoindentation using Atomic Force Microscopy, and their elastic modulus was found to be approximately 17.2 ± 1 MPa. At a cycle of piezo expansion-retraction (loading-unloading) of a silicon tip on a fiber, relatively high adhesion was observed during unloading. It is proposed that this was primarily due to molecular rearrangements at the utmost layers of the fiber caused by the indentation of the hydrophilic tip. The phospholipid fibers were shown to be stable in ambient conditions, preserving the modulus of elasticity up to 24 h.

Nanomechanics of electrospun phospholipid fiber

DEFF Research Database (Denmark)

Mendes, Ana Carina Loureiro; Nikogeorgos, Nikolaos; Lee, Seunghwan

2015-01-01

Electrospun asolectin phospholipid fibers were prepared using isooctane as a solvent and had an average diameter of 6.1 +/- 2.7 mu m. Their mechanical properties were evaluated by nanoindentation using Atomic Force Microscopy, and their elastic modulus was found to be approximately 17.2 +/- 1MPa....... At a cycle of piezo expansion-retraction (loading-unloading) of a silicon tip on a fiber, relatively high adhesion was observed during unloading. It is proposed that this was primarily due to molecular rearrangements at the utmost layers of the fiber caused by the indentation of the hydrophilic tip....... The phospholipid fibers were shown to be stable in ambient conditions, preserving the modulus of elasticity up to 24 h. (c) 2015 AIP Publishing LLC....

The dynamics of the G protein-coupled neuropeptide Y2 receptor in monounsaturated membranes investigated by solid-state NMR spectroscopy

Energy Technology Data Exchange (ETDEWEB)

Thomas, Lars; Kahr, Julian; Schmidt, Peter; Krug, Ulrike; Scheidt, Holger A.; Huster, Daniel, E-mail: daniel.huster@medizin.uni-leipzig.de [University of Leipzig, Institute of Medical Physics and Biophysics (Germany)

2015-04-15

In contrast to the static snapshots provided by protein crystallography, G protein-coupled receptors constitute a group of proteins with highly dynamic properties, which are required in the receptors’ function as signaling molecule. Here, the human neuropeptide Y2 receptor was reconstituted into a model membrane composed of monounsaturated phospholipids and solid-state NMR was used to characterize its dynamics. Qualitative static {sup 15}N NMR spectra and quantitative determination of {sup 1}H–{sup 13}C order parameters through measurement of the {sup 1}H–{sup 13}C dipolar couplings of the CH, CH{sub 2} and CH{sub 3} groups revealed axially symmetric motions of the whole molecule in the membrane and molecular fluctuations of varying amplitude from all molecular segments. The molecular order parameters (S{sub backbone} = 0.59–0.67, S{sub CH2} = 0.41–0.51 and S{sub CH3} = 0.22) obtained in directly polarized {sup 13}C NMR experiments demonstrate that the Y2 receptor is highly mobile in the native-like membrane. Interestingly, according to these results the receptor was found to be slightly more rigid in the membranes formed by the monounsaturated phospholipids than by saturated phospholipids as investigated previously. This could be caused by an increased chain length of the monounsaturated lipids, which may result in a higher helical content of the receptor. Furthermore, the incorporation of cholesterol, phosphatidylethanolamine, or negatively charged phosphatidylserine into the membrane did not have a significant influence on the molecular mobility of the Y2 receptor.

Fatty acid composition of total lipids and phospholipids of muscular tissue and brain of rats under the impact of vibration

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N. M. Kostyshyn

2016-06-01

Full Text Available Fatty acids are important structural components of biological membranes, energy substrate of cells involved in fixing phospholipid bilayer proteins, and acting as regulators and modulators of enzymatic activity. Under the impact of vibration oscillations there can occur shifts in the ratio of different groups of fatty acids, and degrees of their saturation may change. The imbalance between saturated, monounsaturated and polyunsaturated fatty acids, which occurs later in the cell wall, disrupts fluidity and viscosity of lipid phase and causes abnormal cellular metabolism. Aim. In order to study the impact of vibration on the level of fatty acids of total lipids in muscular tissue and fatty acid composition of phospholipids in muscles and brain, experimental animals have been exposed to vertical vibration oscillations with different frequency for 28 days. Methods and results. Tissues fragments of hip quadriceps and brain of rats were used for obtaining methyl esters of fatty acids studied by the method of gas-liquid chromatography. It was found that the lipid content, ratio of its separate factions and fatty acid composition in muscular tissue and brain of animals with the action of vibration considerably varies. With the increase of vibration acceleration tendency to increase in absolute quantity of total lipids fatty acids can be observed at the account of increased level of saturated and monounsaturated ones. These processes are caused by activation of self-defense mechanisms of the body under the conditions of deviations from stabilized physiological norm, since adaptation requires certain structural and energy costs. Increase in the relative quantity of saturated and monounsaturated fatty acids in phospholipids of muscles and brain and simultaneous reduction in concentration of polyunsaturated fatty acids are observed. Conclusion. These changes indicate worsening of structural and functional organization of muscles and brain cell membranes of

Adsorption of GST-PI3Kγ at the Air-Buffer Interface and at Substrate and Nonsubstrate Phospholipid Monolayers

Science.gov (United States)

Hermelink, Antje; Kirsch, Cornelia; Klinger, Reinhard; Reiter, Gerald; Brezesinski, Gerald

2009-01-01

The recruitment of phosphoinositide 3-kinase γ (PI3Kγ) to the cell membrane is a crucial requirement for the initiation of inflammation cascades by second-messenger production. In addition to identifying other regulation pathways, it has been found that PI3Kγ is able to bind phospholipids directly. In this study, the adsorption behavior of glutathione S-transferase (GST)-PI3Kγ to nonsubstrate model phospholipids, as well as to commercially available substrate inositol phospholipids (phosphoinositides), was investigated by use of infrared reflection-absorption spectroscopy (IRRAS). The nonsubstrate phospholipid monolayers also yielded important information about structural requirements for protein adsorption. The enzyme did not interact with condensed zwitterionic or anionic monolayers; however, it could penetrate into uncompressed fluid monolayers. Compression to values above its equilibrium pressure led to a squeezing out and desorption of the protein. Protein affinity for the monolayer surface increased considerably when the lipid had an anionic headgroup and contained an arachidonoyl fatty acyl chain in sn-2 position. Similar results on a much higher level were observed with substrate phosphoinositides. No structural response of GST-PI3Kγ to lipid interaction was detected by IRRAS. On the other hand, protein adsorption caused a condensing effect in phosphoinositide monolayers. In addition, the protein reduced the charge density at the interface probably by shifting the pK values of the phosphate groups attached to the inositol headgroups. Because of their strongly polar headgroups, an interaction of the inositides with the water molecules of the subphase can be expected. This interaction is disturbed by protein adsorption, causing the ionization state of the phosphates to change. PMID:19186139

Adsorption of GST-PI3Kgamma at the air-buffer interface and at substrate and nonsubstrate phospholipid monolayers.

Science.gov (United States)

Hermelink, Antje; Kirsch, Cornelia; Klinger, Reinhard; Reiter, Gerald; Brezesinski, Gerald

2009-02-01

The recruitment of phosphoinositide 3-kinase gamma (PI3Kgamma) to the cell membrane is a crucial requirement for the initiation of inflammation cascades by second-messenger production. In addition to identifying other regulation pathways, it has been found that PI3Kgamma is able to bind phospholipids directly. In this study, the adsorption behavior of glutathione S-transferase (GST)-PI3Kgamma to nonsubstrate model phospholipids, as well as to commercially available substrate inositol phospholipids (phosphoinositides), was investigated by use of infrared reflection-absorption spectroscopy (IRRAS). The nonsubstrate phospholipid monolayers also yielded important information about structural requirements for protein adsorption. The enzyme did not interact with condensed zwitterionic or anionic monolayers; however, it could penetrate into uncompressed fluid monolayers. Compression to values above its equilibrium pressure led to a squeezing out and desorption of the protein. Protein affinity for the monolayer surface increased considerably when the lipid had an anionic headgroup and contained an arachidonoyl fatty acyl chain in sn-2 position. Similar results on a much higher level were observed with substrate phosphoinositides. No structural response of GST-PI3Kgamma to lipid interaction was detected by IRRAS. On the other hand, protein adsorption caused a condensing effect in phosphoinositide monolayers. In addition, the protein reduced the charge density at the interface probably by shifting the pK values of the phosphate groups attached to the inositol headgroups. Because of their strongly polar headgroups, an interaction of the inositides with the water molecules of the subphase can be expected. This interaction is disturbed by protein adsorption, causing the ionization state of the phosphates to change.

Interaction of a novel antimicrobial peptide isolated from the venom of solitary bee Colletes daviesanus with phospholipid vesicles and Escherichia coli cells.

Science.gov (United States)

Čujová, Sabína; Bednárová, Lucie; Slaninová, Jiřina; Straka, Jakub; Čeřovský, Václav

2014-11-01

The peptide named codesane (COD), consisting of 18 amino acid residues and isolated from the venom of wild bee Colletes daviesanus (Hymenoptera : Colletidae), falls into the category of cationic α-helical amphipathic antimicrobial peptides. In our investigations, synthetic COD exhibited antimicrobial activity against Gram-positive and Gram-negative bacteria and Candida albicans but also noticeable hemolytic activity. COD and its analogs (collectively referred to as CODs) were studied for the mechanism of their action. The interaction of CODs with liposomes led to significant leakage of calcein entrapped in bacterial membrane-mimicking large unilamellar vesicles made preferentially from anionic phospholipids while no calcein leakage was observed from zwitterionic liposomes mimicking membranes of erythrocytes. The preference of CODs for anionic phospholipids was also established by the blue shift in the tryptophan emission spectra maxima when the interactions of tryptophan-containing COD analogs with liposomes were examined. Those results were in agreement with the antimicrobial and hemolytic activities of CODs. Moreover, we found that the studied peptides permeated both the outer and inner cytoplasmic membranes of Escherichia coli. This was determined by measuring changes in the fluorescence of probe N-phenyl-1-naphthylamine and detecting cytoplasmic β-galactosidase released during the interaction of peptides with E. coli cells. Transmission electron microscopy revealed that treatment of E. coli with one of the COD analogs caused leakage of bacterial content mainly from the septal areas of the cells. Copyright © 2014 European Peptide Society and John Wiley & Sons, Ltd.

Growth and instability of a phospholipid vesicle in a bath of fatty acids

Science.gov (United States)

Dervaux, J.; Noireaux, V.; Libchaber, A. J.

2017-06-01

Using a microfluidic trap, we study the behavior of individual phospholipid vesicles in contact with fatty acids. We show that spontaneous fatty acids insertion inside the bilayer is controlled by the vesicle size, osmotic pressure difference across the membrane and fatty acids concentration in the external bath. Depending on these parameters, vesicles can grow spherically or become unstable and fragment into several daughter vesicles. We establish the phase diagram for vesicle growth and we derive a simple thermodynamic model that reproduces the time evolution of the vesicle volume. Finally, we show that stable growth can be achieved on an artificial cell expressing a simple set of bacterial cytoskeletal proteins, paving the way toward artificial cell reproduction.

Apoptotic Bax at Oxidatively Stressed Mitochondrial Membranes: Lipid Dynamics and Permeabilization

Czech Academy of Sciences Publication Activity Database

Dilgendein, A. P.; Pokorná, Šárka; Lidman, M.; Sparrman, T.; Šachl, Radek; Hof, Martin; Gröbner, G.

2017-01-01

Roč. 112, č. 10 (2017), s. 2147-2158 ISSN 0006-3495 R&D Projects: GA ČR(CZ) GBP208/12/G016 Institutional support: RVO:61388955 Keywords : LOW-FREQUENCY MOTION * OXIDIZED PHOSPHOLIPIDS * BILAYER-MEMBRANES Subject RIV: CF - Physical ; Theoretical Chemistry OBOR OECD: Physical chemistry Impact factor: 3.656, year: 2016

Phospholipid alterations in cardiac sarcoplasmic reticulum induced by xanthine oxidase: contamination of commercial preparations of xanthine oxidase by phospholipase A2

International Nuclear Information System (INIS)

Gamache, D.A.; Kornberg, L.J.; Bartolf, M.; Franson, R.C.

1986-01-01

Incubation of cardiac sarcoplasmic reticulum with xanthine oxidase alone at pH 7.0 resulted in a loss of lipid phosphorus that was potentiated by the addition of xanthine. Using autoclaved E.coli with 1- 14 C-oleate in the 2-acyl position of membrane phospholipids, the authors demonstrate that many, but not all, commercial preparations of xanthine oxidase contain significant phospholipase A 2 (PLA 2 ) activity (64.3-545.6 nmols/min/mg). The PLA 2 was maximally active in the neutral-alkaline pH range, was Ca 2+ -dependent, and was unaffected by the addition of xanthine. PLA 2 activity was totally inhibited by 1mM EDTA whereas radical production by optimal concentrations of xanthine/xanthine oxidase (X/XO) was unaffected by EDTA. Chromatographically purified xanthine oxidase (Sigma Grade III) contained high levels of PLA 2 activity (64.3 nmols/min/mg) compared to endogenous levels of neutral-active, Ca 2+ -dependent PLA 2 measured in various tissue homogenates (≤ 0.5 nmols/ min/mg). Because X/XO mixtures are used extensively to study oxygen free radical-induced cell injury and membrane phospholipid alterations, the presence of a potent extracellular PLA 2 may have influenced previously published reports, and such studies should be interpreted cautiously

Double-chain phospholipid end-capped polyurethanes: Synthesis, characterization and platelet adhesion study

International Nuclear Information System (INIS)

Tan Dongsheng; Zhang Xiaoqing; Li Jiehua; Tan Hong; Fu Qiang

2012-01-01

A novel phospholipid containing double chains and phosphotidylcholine polar head groups, 2-(10-(2-aminoethylamino)-10-oxodecanamido)-3-(decyloxy)-3-oxopropyl phosphorylcholine (ADDPC), was synthesized and characterized. Two kinds of double-chain phospholipid end-capped polyurethanes with different soft segments were prepared. The structure of prepared polyurethanes was characterized by X-ray photoelectron spectroscopic (XPS), attenuated total reflection Fourier transform infrared (ATR-FTIR) spectrometry and atomic force microscope (AFM), which indicated that the double-chain phospholipids enriched onto the top surface of the prepared polyurethane films. The preliminary evaluation of blood compatibility showed that these novel phospholipid end-capped polyurethanes could suppress platelet adhesion and activation effectively. This property did not depend on the chemical structure of polyurethanes. In addition, according to tensile test results, the phospholipid polyurethanes kept good mechanical properties in comparison with original polyurethanes. It is suggested that double-chain phospholipid end-caption has good potential for achieving both hemocompatibility and good mechanical properties simultaneously for polyurethanes.

Novel Blend Membranes Based on Acid-Base Interactions for Fuel Cells

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Yongzhu Fu

2012-10-01

Full Text Available Fuel cells hold great promise for wide applications in portable, residential, and large-scale power supplies. For low temperature fuel cells, such as the proton exchange membrane fuel cells (PEMFCs and direct methanol fuel cells (DMFCs, proton-exchange membranes (PEMs are a key component determining the fuel cells performance. PEMs with high proton conductivity under anhydrous conditions can allow PEMFCs to be operated above 100 °C, enabling use of hydrogen fuels with high-CO contents and improving the electrocatalytic activity. PEMs with high proton conductivity and low methanol crossover are critical for lowering catalyst loadings at the cathode and improving the performance and long-term stability of DMFCs. This review provides a summary of a number of novel acid-base blend membranes consisting of an acidic polymer and a basic compound containing N-heterocycle groups, which are promising for PEMFCs and DMFCs.

The solubilisation of boar sperm membranes by different detergents - a microscopic, MALDI-TOF MS, 31P NMR and PAGE study on membrane lysis, extraction efficiency, lipid and protein composition

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Müller Karin

2009-11-01

Full Text Available Abstract Background Detergents are often used to isolate proteins, lipids as well as "detergent-resistant membrane domains" (DRMs from cells. Different detergents affect different membrane structures according to their physico-chemical properties. However, the effects of different detergents on membrane lysis of boar spermatozoa and the lipid composition of DRMs prepared from the affected sperm membranes have not been investigated so far. Results Spermatozoa were treated with the selected detergents Pluronic F-127, sodium cholate, CHAPS, Tween 20, Triton X-100 and Brij 96V. Different patterns of membrane disintegration were observed by light and electron microscopy. In accordance with microscopic data, different amounts of lipids and proteins were released from the cells by the different detergents. The biochemical methods to assay the phosphorus and cholesterol contents as well as 31P NMR to determine the phospholipids were not influenced by the presence of detergents since comparable amounts of lipids were detected in the organic extracts from whole cell suspensions after exposure to each detergent. However, matrix-assisted laser desorption and ionization time-of-flight mass spectrometry applied to identify phospholipids was essentially disturbed by the presence of detergents which exerted particular suppression effects on signal intensities. After separation of the membrane fractions released by detergents on a sucrose gradient only Triton X-100 and sodium cholate produced sharp turbid DRM bands. Only membrane solubilisation by Triton X-100 leads to an enrichment of cholesterol, sphingomyelin, phosphatidylinositol and phosphatidylethanolamine in a visible DRM band accompanied by a selective accumulation of proteins. Conclusion The boar sperm membranes are solubilised to a different extent by the used detergents. Particularly, the very unique DRMs isolated after Triton X-100 exposure are interesting candidates for further studies regarding the

Phosphatidylinositol 3-phosphates-at the interface between cell signalling and membrane traffic.

Science.gov (United States)

Marat, Andrea L; Haucke, Volker

2016-03-15

Phosphoinositides (PIs) form a minor class of phospholipids with crucial functions in cell physiology, ranging from cell signalling and motility to a role as signposts of compartmental membrane identity. Phosphatidylinositol 3-phosphates are present at the plasma membrane and within the endolysosomal system, where they serve as key regulators of both cell signalling and of intracellular membrane traffic. Here, we provide an overview of the metabolic pathways that regulate cellular synthesis of PI 3-phosphates at distinct intracellular sites and discuss the mechanisms by which these lipids regulate cell signalling and membrane traffic. Finally, we provide a framework for how PI 3-phosphate metabolism is integrated into the cellular network. © 2016 The Authors.

Recent Advances in Phospholipids from Colostrum, Milk and Dairy By-Products

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Vito Verardo

2017-01-01

Full Text Available Milk is one of the most important foods for mammals, because it is the first form of feed providing energy, nutrients and immunological factors. In the last few years, milk lipids have attracted the attention of researchers due to the presence of several bioactive components in the lipid fraction. The lipid fraction of milk and dairy products contains several components of nutritional significance, such as ω-3 and ω-6 polyunsaturated fatty acids, CLA, short chain fatty acids, gangliosides and phospholipids. Prospective cohort evidence has shown that phospholipids play an important role in the human diet and reinforce the possible relationship between their consumption and prevention of several chronic diseases. Because of these potential benefits of phospholipids in the human diet, this review is focused on the recent advances in phospholipids from colostrum, milk and dairy by-products. Phospholipid composition, its main determination methods and the health activities of these compounds will be addressed.

Exogenous Polyunsaturated Fatty Acids Impact Membrane Remodeling and Affect Virulence Phenotypes among Pathogenic Vibrio Species.

Science.gov (United States)

Moravec, Anna R; Siv, Andrew W; Hobby, Chelsea R; Lindsay, Emily N; Norbash, Layla V; Shults, Daniel J; Symes, Steven J K; Giles, David K

2017-11-15

The pathogenic Vibrio species ( V. cholerae , V. parahaemolyticus , and V. vulnificus ) represent a constant threat to human health, causing foodborne and skin wound infections as a result of ingestion of or exposure to contaminated water and seafood. Recent studies have highlighted Vibrio 's ability to acquire fatty acids from environmental sources and assimilate them into cell membranes. The possession and conservation of such machinery provokes consideration of fatty acids as important factors in the pathogenic lifestyle of Vibrio species. The findings here link exogenous fatty acid exposure to changes in bacterial membrane phospholipid structure, permeability, phenotypes associated with virulence, and consequent stress responses that may impact survival and persistence of pathogenic Vibrio species. Polyunsaturated fatty acids (PUFAs) (ranging in carbon length and unsaturation) supplied in growth medium were assimilated into bacterial phospholipids, as determined by thin-layer chromatography and liquid chromatography-mass spectrometry. The incorporation of fatty acids variably affected membrane permeability, as judged by uptake of the hydrophobic compound crystal violet. For each species, certain fatty acids were identified as affecting resistance to antimicrobial peptide treatment. Significant fluctuations were observed with regard to both motility and biofilm formation following growth in the presence of individual PUFAs. Our results illustrate the important and complex roles of exogenous fatty acids in the membrane physiology and virulence of a bacterial genus that inhabits aquatic and host environments containing an abundance of diverse fatty acids. IMPORTANCE Bacterial responses to fatty acids include, but are not limited to, degradation for metabolic gain, modification of membrane lipids, alteration of protein function, and regulation of gene expression. Vibrio species exhibit significant diversity with regard to the machinery known to participate in the

Self-assembling peptides form nanodiscs that stabilize membrane proteins

DEFF Research Database (Denmark)

Midtgaard, Søren Roi; Pedersen, Martin Cramer; Kirkensgaard, Jacob Judas Kain

2014-01-01

-ray scattering (SAXS) and small-angle neutron scattering (SANS) supported by coarse-grained molecular dynamics simulations. The detailed structure of the discs was determined in unprecedented detail and it was found that they adopt a discoidal structure very similar to the ApoA1 based nanodiscs. We furthermore...... show that, like the ApoA1 and derived nanodiscs, these peptide discs can accommodate and stabilize a membrane protein. Finally, we exploit their dynamic properties and show that the 18A discs may be used for transferring membrane proteins and associated phospholipids directly and gently......New methods to handle membrane bound proteins, e.g. G-protein coupled receptors (GPCRs), are highly desirable. Recently, apoliprotein A1 (ApoA1) based lipoprotein particles have emerged as a new platform for studying membrane proteins, and it has been shown that they can self...

Quantitative studies of antimicrobial peptide-lipid membrane interactions

DEFF Research Database (Denmark)

Kristensen, Kasper

antimicrobial peptides interact with phospholipid membranes. Motivated by that fact, the scope of this thesis is to study these antimicrobial peptide-lipid membrane interactions. In particular, we attempt to study these interactions with a quantitative approach. For that purpose, we consider the three...... a significant problem for quantitative studies of antimicrobial peptide-lipid membrane interactions; namely that antimicrobial peptides adsorb to surfaces of glass and plastic. Specifically, we demonstrate that under standard experimental conditions, this effect is significant for mastoparan X, melittin...... lead to inaccurate conclusions, or even completely wrong conclusions, when interpreting the FCS data. We show that, if all of the pitfalls are avoided, then FCS is a technique with a large potential for quantitative studies of antimicrobial peptide-induced leakage of fluorescent markers from large...

Vapor-phase etching of InP using anhydrous HCl and PH/sub 3/ gas

International Nuclear Information System (INIS)

Pak, K.; Koide, Y.; Imai, K.; Yoshida, A.; Nakamura, T.; Yasuda, Y.; Nishinaga, T.

1986-01-01

In situ etching of the substrate surface for vapor-phase epitaxy is a useful technique for obtaining a smooth and damage-free surface prior to the growth. Previous work showed that the incorporation of in situ etching of InP substrate with anhydrous HCl gas resulted in a significant improvement in the surface morphologies for MOVPE-grown InGaAs/InP and InP epitaxial layers. However, the experiment on the HCl etching of the InP substrate for a wide temperature range has not been performed as yet. In this note, the authors describe the effect of the substrate temperature on the etching morphology of InP substrate by using the anhydrous HCl and PH/sub 3/ gases. In the experiment, they used a standard MOVPE horizontal system. A quartz reactor tube in a 60 mm ID, 60 cm long, was employed

The structural role of cholesterol in cell membranes: from condensed bilayers to lipid rafts.

Science.gov (United States)

Krause, Martin R; Regen, Steven L

2014-12-16

proposed for cholesterol's condensing effect: (i) an umbrella mechanism in which the acyl chains and cholesterol become more tightly packed as cholesterol content increases because they share limited space under phospholipid headgroups and (ii) a template mechanism whereby cholesterol functions as a planar hydrophobic template at the membrane surface, thereby maximizing hydrophobic interactions and the hydrophobic effect. Specifically, our NNR experiments rule out the umbrella mechanism and provide strong support for the template mechanism. Similar NNR measurements have also allowed us to address the question of whether the interactions between low-melting kinked phospholipids and cholesterol can play a significant role in the formation of lipid rafts. Specifically, these NNR measurements have led to our discovery of a new physical principle in the lipids and membranes area that must be operating in biological membranes, that is, a "push-pull" mechanism, whereby cholesterol is pushed away from low-melting phospholipids and pulled toward high-melting lipids. Thus, to the extent that lipid rafts play a role in the functioning of cell membranes, low-melting phospholipids must be active participants.

Crystallo-chemistry of boric anhydride and of anhydrous borates

International Nuclear Information System (INIS)

Bernardin, Jacques

1968-01-01

After an overview of various aspects related to the atomic structure of boron and of its three-bind and four-bind compounds, this report briefly presents the different forms of boric anhydride (in solid, liquid, glassy and gaseous state), presents and comments the structure of these different forms, and addresses the molten boric anhydride which is used as oxide solvent. The next part addresses the structure of anhydrous borates. It presents some generalities on their structure, and describes examples of known structures: dimers, trimers, polymers with a degree higher than three like calcium metaborate, caesium tri-borate, lithium tetraborate, or potassium pentaborate

Using crosslinkable diacetylene phospholipids to construct two-dimensional packed beds in supported lipid bilayer separation platforms

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Shu-Kai Hu, Sheng-Wen Hsiao, Hsun-Yen Mao, Ya-Ming Chen, Yung Chang and Ling Chao

2013-01-01

Full Text Available Separating and purifying cell membrane-associated biomolecules has been a challenge owing to their amphiphilic property. Taking these species out of their native lipid membrane environment usually results in biomolecule degradation. One of the new directions is to use supported lipid bilayer (SLB platforms to separate the membrane species while they are protected in their native environment. Here we used a type of crosslinkable diacetylene phospholipids, diynePC (1,2-bis(10,12-tricosadiynoyl-sn-glycero-3-phosphocholine, as a packed material to create a 'two-dimensional (2D packed bed' in a SLB platform. After the diynePC SLB is exposed to UV light, some of the diynePC lipids in the SLB can crosslink and the non-crosslinked monomer lipids can be washed away, leaving a 2D porous solid matrix. We incorporated the lipid vesicle deposition method with a microfluidic device to pattern the location of the packed-bed region and the feed region with species to be separated in a SLB platform. Our atomic force microscopy result shows that the nano-scaled structure density of the '2D packed bed' can be tuned by the UV dose applied to the diynePC membrane. When the model membrane biomolecules were forced to transport through the packed-bed region, their concentration front velocities were found to decrease linearly with the UV dose, indicating the successful creation of packed obstacles in these 2D lipid membrane separation platforms.

Interaction of Local Anesthetics with Biomembranes Consisting of Phospholipids and Cholesterol: Mechanistic and Clinical Implications for Anesthetic and Cardiotoxic Effects

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Hironori Tsuchiya

2013-01-01

Full Text Available Despite a long history in medical and dental application, the molecular mechanism and precise site of action are still arguable for local anesthetics. Their effects are considered to be induced by acting on functional proteins, on membrane lipids, or on both. Local anesthetics primarily interact with sodium channels embedded in cell membranes to reduce the excitability of nerve cells and cardiomyocytes or produce a malfunction of the cardiovascular system. However, the membrane protein-interacting theory cannot explain all of the pharmacological and toxicological features of local anesthetics. The administered drug molecules must diffuse through the lipid barriers of nerve sheaths and penetrate into or across the lipid bilayers of cell membranes to reach the acting site on transmembrane proteins. Amphiphilic local anesthetics interact hydrophobically and electrostatically with lipid bilayers and modify their physicochemical property, with the direct inhibition of membrane functions, and with the resultant alteration of the membrane lipid environments surrounding transmembrane proteins and the subsequent protein conformational change, leading to the inhibition of channel functions. We review recent studies on the interaction of local anesthetics with biomembranes consisting of phospholipids and cholesterol. Understanding the membrane interactivity of local anesthetics would provide novel insights into their anesthetic and cardiotoxic effects.

Charge asymmetry of the purple membrane measured by uranyl quenching of dansyl fluorescence. [Halobacterium halobium

Energy Technology Data Exchange (ETDEWEB)

Renthal, R.; Cha, C.H.

1984-05-01

Purple membrane was covalently labeled with 5-(dimethylamino) naphthalene-1-sulfonyl hydrazine (dansyl hydrazine) by carbodiimide coupling to the cytoplasmic surface (carboxyl-terminal tail: 0.7 mol/mol bacteriorhodopsin) or by periodate oxidation and dimethylaminoborane reduction at the extracellular surface (glycolipids: 1 mol/mol). In 2 mM acetate buffer, pH 5.6, micromolar concentrations of UO/sub 2//sup 2 +/ were found to quench the dansyl groups on the cytoplasmic surface (maximum = 26%), while little quenching was observed at the extracellular surface (maximum = 4%). Uranyl ion quenched dansyl hydrazine in free solution at much higher concentrations. Uranyl also bound tightly to unmodified purple membrane, (apparent dissociation constant = 0.8 ..mu..M) as measured by a centrifugation assay. The maximum stoichiometry was 10 mol/mol of bacteriorhodopsin, which is close to the amount of phospholipid phosphorus in purple membrane. The results were analyzed on the assumptions that UO/sub 2//sup 2 +/ binds in a 1:1 complex with phospholipid phosphate and that the dansyl distributon and quenching mechanisms are the same at both surfaces. This indicates a 9:1 ratio of phosphate between the cytoplasmic and extracellular surfaces. Thus, the surface change density of the cytoplasmic side of the membrane is more negative than - 0.010 charges/A/sup 2/.

Hybrid electrospun chitosan-phospholipids nanofibers for transdermal drug delivery

DEFF Research Database (Denmark)

Mendes, Ana Carina Loureiro; Gorzelanny, Christian; Halter, Natalia

2016-01-01

Chitosan (Ch) polysaccharide was mixed with phospholipids (P) to generate electrospun hybrid nanofibers intended to be used as platforms for transdermal drug delivery. Ch/P nanofibers exibithed average diameters ranging from 248 +/- 94 nm to 600 +/- 201 nm, depending on the amount of phospholipids...... used. Fourier Transformed Infra-Red (FTIR) spectroscopy and Dynamic Light Scattering (DLS) data suggested the occurrence of electrostatic interactions between amine groups of chitosan with the phospholipid counterparts. The nanofibers were shown to be stable for at least 7 days in Phosphate Buffer...... culture plate (control). The release of curcumin, diclofenac and vitamin B12, as model drugs, from Ch/P hybrid nanofibers was investigated, demonstrating their potential utilization as a transdermal drug delivery system....

Alboserpin, a Factor Xa Inhibitor from the Mosquito Vector of Yellow Fever, Binds Heparin and Membrane Phospholipids and Exhibits Antithrombotic Activity

Czech Academy of Sciences Publication Activity Database

Calvo, E.; Mizurini, D.M.; Sa-Nunes, A.; Ribeiro, J.M.C.; Andersen, J. F.; Mans, B.J.; Monteiro, R.Q.; Kotsyfakis, Michalis; Francischetti, I.M.B.

2011-01-01

Roč. 286, č. 32 (2011), 27998-28010 ISSN 0021-9258 Institutional research plan: CEZ:AV0Z60220518 Keywords : serpin * mosquito * Aedes albopictus * phospholipids * Factor Xa * heparin * binding affinity * coagulation * thrombus * bleeding Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 4.773, year: 2011

Membrane fusion and exocytosis.

Science.gov (United States)

Jahn, R; Südhof, T C

1999-01-01

Membrane fusion involves the merger of two phospholipid bilayers in an aqueous environment. In artificial lipid bilayers, fusion proceeds by means of defined transition states, including hourglass-shaped intermediates in which the proximal leaflets of the fusing membranes are merged whereas the distal leaflets are separate (fusion stalk), followed by the reversible opening of small aqueous fusion pores. Fusion of biological membranes requires the action of specific fusion proteins. Best understood are the viral fusion proteins that are responsible for merging the viral with the host cell membrane during infection. These proteins undergo spontaneous and dramatic conformational changes upon activation. In the case of the paradigmatic fusion proteins of the influenza virus and of the human immunodeficiency virus, an amphiphilic fusion peptide is inserted into the target membrane. The protein then reorients itself, thus forcing the fusing membranes together and inducing lipid mixing. Fusion of intracellular membranes in eukaryotic cells involves several protein families including SNAREs, Rab proteins, and Sec1/Munc-18 related proteins (SM-proteins). SNAREs form a novel superfamily of small and mostly membrane-anchored proteins that share a common motif of about 60 amino acids (SNARE motif). SNAREs reversibly assemble into tightly packed helical bundles, the core complexes. Assembly is thought to pull the fusing membranes closely together, thus inducing fusion. SM-proteins comprise a family of soluble proteins that bind to certain types of SNAREs and prevent the formation of core complexes. Rab proteins are GTPases that undergo highly regulated GTP-GDP cycles. In their GTP form, they interact with specific proteins, the effector proteins. Recent evidence suggests that Rab proteins function in the initial membrane contact connecting the fusing membranes but are not involved in the fusion reaction itself.

REDOR NMR Reveals Multiple Conformers for a Protein Kinase C Ligand in a Membrane Environment

Directory of Open Access Journals (Sweden)

Hao Yang

2018-01-01

Full Text Available Bryostatin 1 (henceforth bryostatin is in clinical trials for the treatment of Alzheimer’s disease and for HIV/AIDS eradication. It is also a preclinical lead for cancer immunotherapy and other therapeutic indications. Yet nothing is known about the conformation of bryostatin bound to its protein kinase C (PKC target in a membrane microenvironment. As a result, efforts to design more efficacious, better tolerated, or more synthetically accessible ligands have been limited to structures that do not include PKC or membrane effects known to influence PKC–ligand binding. This problem extends more generally to many membrane-associated proteins in the human proteome. Here, we use rotational-echo double-resonance (REDOR solid-state NMR to determine the conformations of PKC modulators bound to the PKCδ-C1b domain in the presence of phospholipid vesicles. The conformationally limited PKC modulator phorbol diacetate (PDAc is used as an initial test substrate. While unanticipated partitioning of PDAc between an immobilized protein-bound state and a mobile state in the phospholipid assembly was observed, a single conformation in the bound state was identified. In striking contrast, a bryostatin analogue (bryolog was found to exist exclusively in a protein-bound state, but adopts a distribution of conformations as defined by three independent distance measurements. The detection of multiple PKCδ-C1b-bound bryolog conformers in a functionally relevant phospholipid complex reveals the inherent dynamic nature of cellular systems that is not captured with single-conformation static structures. These results indicate that binding, selectivity, and function of PKC modulators, as well as the design of new modulators, are best addressed using a dynamic multistate model, an analysis potentially applicable to other membrane-associated proteins.

Electrolysis of a nitrosyl hexafluoro-molybdate in anhydrous hydrogen fluoride

International Nuclear Information System (INIS)

Mougin, Jacques

1972-01-01

This thesis addresses the field of irradiated fuel reprocessing, and more particularly the study of the formation of molybdenum hexafluoride (MoF_6) by electrolysis of nitrosyl hexafluoro-molybdate (NOMoF_6) in solution in HF. The author presents the conditions of preparation of an electrolysis in anhydrous HF: solvent purification and control, production of a reference electrode, potential-kinetic study of the behaviour of materials selected for the electrode. The author then addresses the actual hydrolysis of the solution of nitrosyl hexafluoro-molybdate [fr

Yeast-expressed human membrane protein aquaporin-1 yields excellent resolution of solid-state MAS NMR spectra

International Nuclear Information System (INIS)

Emami, Sanaz; Fan Ying; Munro, Rachel; Ladizhansky, Vladimir; Brown, Leonid S.

2013-01-01

One of the biggest challenges in solid-state NMR studies of membrane proteins is to obtain a homogeneous natively folded sample giving high spectral resolution sufficient for structural studies. Eukaryotic membrane proteins are especially difficult and expensive targets in this respect. Methylotrophic yeast Pichia pastoris is a reliable producer of eukaryotic membrane proteins for crystallography and a promising economical source of isotopically labeled proteins for NMR. We show that eukaryotic membrane protein human aquaporin 1 can be doubly ( 13 C/ 15 N) isotopically labeled in this system and functionally reconstituted into phospholipids, giving excellent resolution of solid-state magic angle spinning NMR spectra.

Efficient discrimination and removal of phospholipids during electromembrane extraction from human plasma samples

DEFF Research Database (Denmark)

Vårdal, Linda; Gjelstad, Astrid; Huang, Chuixiu

2017-01-01

to be highly efficient for providing phospholipid-free extracts. CONCLUSION: Ultra-HPLC-MS/MS analysis of the donor solutions revealed that the phospholipids principally remained in the plasma samples. This proved that the phospholipids did not migrate in the electrical field and they were prevented from......AIM: For the first time, extracts obtained from human plasma samples by electromembrane extraction (EME) were investigated comprehensively with particular respect to phospholipids using ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). Thhe purpose...

The chloroplasts membrane phospholipids of Chlamydomonas reinhardii mutant not forming the Photosystem 2

International Nuclear Information System (INIS)

Trusova, V.M.; Ladygin, V.G.; Mezentsev, V.V.; Molchanov, M.I.

1987-01-01

Study on a component composition and physical state of photosynthetic membranes of Chlamydomonas chloroplasts of the wild type and mutant A-110 with disturbance of electron transfer chain in the photosystem 2 region permitted to conclude that 170 A diameter particles localized on the internal hydrophobic surface of membrane chips are deleted with respect to phosphatidylglycerin. The results obtained permit to suggest that the formation of protein-lipid complexes containing phosphatidylglycerins is suppressed in mutant A-110 which is not capable of the lamellar system differentation in

Interactions of Borneol with DPPC Phospholipid Membranes: A Molecular Dynamics Simulation Study

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Qianqian Yin

2014-11-01

Full Text Available Borneol, known as a “guide” drug in traditional Chinese medicine, is widely used as a natural penetration enhancer in modern clinical applications. Despite a large number of experimental studies on borneol’s penetration enhancing effect, the molecular basis of its action on bio-membranes is still unclear. We carried out a series of coarse-grained molecular dynamics simulations with the borneol concentration ranging from 3.31% to 54.59% (v/v, lipid-free basis to study the interactions of borneol with aDPPC(1,2-dipalmitoylsn-glycero-3-phosphatidylcholine bilayer membrane, and the temperature effects were also considered. At concentrations below 21.89%, borneol’s presence only caused DPPC bilayer thinning and an increase in fluidity; A rise in temperature could promote the diffusing progress of borneol. When the concentration was 21.89% or above, inverted micelle-like structures were formed within the bilayer interior, which led to increased bilayer thickness, and an optimum temperature was found for the interaction of borneol with the DPPC bilayer membrane. These findings revealed that the choice of optimal concentration and temperature is critical for a given application in which borneol is used as a penetration enhancer. Our results not only clarify some molecular basis for borneol’s penetration enhancing effects, but also provide some guidance for the development and applications of new preparations containing borneol.

Plant cell plasma membrane structure and properties under clinostatting

Science.gov (United States)

Polulakh, Yu. A.; Zhadko, S. I.; Klimchuk, D. A.; Baraboy, V. A.; Alpatov, A. N.; Sytnik, K. M.

Structural-functional organization of plasma membrane of pea roots seedling was investigated by methods of chemiluminescence, fluorescence probes, chromatography and freeze-fracture studies under normal conditions and clinostatting. Phase character of lipid peroxidation intensity was fixed. The initial phase of this process is characterized by lipid peroxidation decreasing with its next induction. The primary changes depending on free-radical mechanisms of lipid peroxidation were excellently revealed by chemiluminescence. Plasmalemma microviscosity increased on the average of 15-20 % under microgravity at the initial stages of its phenomenon. There were major changes of phosphatidilcholine and phosphatidilethanolamine contents. The total quantity of phospholipids remained rather stable. Changes of phosphatide acid concentration point to degradation and phospholipids biosynthesis. There were increases of unsaturated fatty acids mainly at the expense of linoleic and linolenic acids and also a decrease of saturated fatty acid content at the expense of palmitic and stearic acids. Unsaturation index of fatty acids increased as well. On the whole fatty acid composition was variable in comparison with phospholipids. Probably it is one of mechanisms of maintaining of microviscosity within definite limits. Considerable structural changes in organization of plasmalemma protein-lipid complex were not revealed by the freeze-fracture studies.

Identification and characterization of polyphosphoinositides in the tonoplast membrane

International Nuclear Information System (INIS)

Cho, Myeon; Memon, A.R.; Boss, W.F.

1989-01-01

In vitro data indicate that the tonoplast membrane is the major target site for IP 3 action and Ca +2 is released from this compartment. If this is true, then several questions arise: (1) Is the plasma membrane or tonoplast the source of IP 3 ? (2) If the plasma membrane is the source of IP 3 , how does IP 3 get to the vacuole? In this connection, the results from in vivo labeling of carrot cells with [ 3 H] inositol show that the major phospholipid components in the tonoplast are PI and lysoPIP. In vitro phosphorylation studies using [γ- 32 P]ATP indicate that PI kinase is present in the tonoplast membrane even though PIP was not detected with inositol labeling. If PIP and PIP 2 are not present in the tonoplast, then IP 3 must diffuse or be transported from the plasma membrane. Since IP 3 is rapidly metabolized by cytosolic phosphatases there may be an IP 3 -sensitive calcium store other than the vacuole closely associated with the plasma membrane

Formation of oriented membrane multilayers of Na/K-ATPase

International Nuclear Information System (INIS)

Pachence, J.M.; Knott, R.; Edelman, I.S.; Schoenborn, B.P.; Wallace, B.A.

1982-01-01

The isolated membrane-bound enzyme retains its ouabain-sensitive ATP hydrolysis activity, and produces ATP-dependent Na + and K + fluxes when incorporated into phospholipid vesicles. The ultimate goal of this work is to determine its low resolution structure using both X-ray and neutron diffraction. A number of methods were used to impart lamellar stacking order to highly purified pig Na/K-ATPase membranes. Upon partial dehydration, x-ray diffraction from Na/K-ATPase membrane multilayers at 98% relative humidity yielded discrete reflections of 118 A periodicity, diffracting to 1/14.8 A -1 , additionally, continuous diffraction to 1/10 A -1 was obtained. Subjecting the membrane multilayers to high magnetic fields improved the quality of the lamellar diffraction dramatically. Neutron diffraction studies of the partially dehydrated Na/K-ATPase membrane multilayers detected a mosaic spread of 2 0 when the samples were subjected to a magnetic field of 5 Tesla perpendicular to the membrane surface; the reflections were narrower than the camera line width; hence, the lattice disorder has also decreased significantly, although only four orders were measured

Mechanics, morphology, and mobility in stratum corneum membranes

Science.gov (United States)

Olmsted, Peter; Das, Chinmay; Noro, Massimo

2012-02-01

The stratum corneum is the outermost layer of skin, and serves as a protective barrier against external agents, and to control moisture. It comprises keratin bodies (corneocytes) embedded in a matrix of lipid bilayers. Unlike the more widely studied phospholipid bilayers, the SC bilayers are typically in a gel-like state. Moreover, the SC membrane composition is radically different from more fluid counterparts: it comprises single tailed fatty acids, ceramides, and cholesterol; with many distinct ceramides possessing different lengths of tails, and always with two tails of different lengths. I will present insight from computer simulations into the morphology, mechanical properties, and diffusion (barrier) properties of these highly heterogeneous membranes. Our results provide some clue as to the design principles for the SC membrane, and is an excellent example of the use of wide polydispersity by natural systems.

Gamma irradiation prolongs the sea bass (Dicentrarchus labrax L) storage and delays the lipids membrane degradation

International Nuclear Information System (INIS)

Barkallah, Insaf; Mahjoub, Abdelmajid; Cheour, F.

2005-01-01

Sea bass have been submitted to gamma rays radiation at doses 0, 1, 2, 3 or 4 KGy at 0,087KGY/min rate and stored during 21 days at 1 0 C to evaluate the effects on the development of microorganisms as well as on the conservation and the degradation of membranes lipids Microbiological analysis has revealed only the presence of mesophils germs in muscles which are totally eliminated at dose 4 KGy. The presence of psychrophiles, halophiles, coliformes, fecaux and totals, Staphylococcus aureus and Clostridium sulfutoreducteurs has been noted in the visceres of sea bass. They are completely eliminated at dose 1KGy. Contents in nitrogen basic fowl (ABVT) and in trimethylamine (TMA), as well as the pH increase during storage of fish in relation with the reduction of phospholipids, the diminution of the degree of insaturation of the phospholipids and the free fat acidic fraction, and the increase of the report sterols on phospholipids. The irradiation to gamma rays allows to slow these changes and consequently reduced the deterioration of the sea bass. We conclude that the irradiation to gamma rays could have prolong the conservation of sea bass sea bass by preserving probably lipids membranes of the degradation and by inhibiting the development of microorganisms

Environmental pollution changes in membrane lipids, antioxidants and vitality of Scots pine (Pinus sylvestris L. pollen

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Paweł M. Pukacki

2011-01-01

Full Text Available Investigations were carried out on pollen grains of Scots pine (Pinus sylvestris L. collected from trees at 1.5, 3, 4 km and control, 20 km from the Luboń factory producing mineral fertilisers. The percentage of germination of pollen formed close to the pollution source was ca 20% lower compared to the control pollen. Lowered vitality of the pollen was effected in changes of the structure of cytoplasmic membranes. Pollen from the polluted area contained ca 15% less total phospholipids, mainly phosphatidylcholine and phosphatytidylinositol and had a lower content of soluble proteins and less of low molecular antioxidants, such as thiols and ascorbic acid. Composition of total fatty acid in phospholipids fractions showed a significant reduction in the degree of unsaturation of fatty acids. Pollen originating from the polluted area and stored at -30°C showed considerably stronger degradation of cytoplasmic membranes than control.

Ultraviolet- and sunlight-induced lipid peroxidation in liposomal membrane

International Nuclear Information System (INIS)

Mandal, T.K.; Chatterjee, S.N.

1980-01-01

Ultraviolet radiation and sunlight caused lipid peroxidation in the liposomal membrane (as detected by measurement of the oxidation index, A 233 /A 215 , and the amount of malondialdehyde formed) and made the membrane leaky (as revealed by the release of the trapped chromate anions). The oxidation index and the formation of malondialdehyde increased linearly with increasing dose of radiation and depended significantly on the dose rate. The effects were smaller in liposomes derived from Vibrio cholerae phospholipid than in those derived from egg lecithin. The effects of the radiation dose and dose rate on hemolysis and peroxidation (MDA formation) of the erythrocyte membrane followed a similar pattern. A direct correlation between the percentage leakage of chromate (Y) and the oxidation index (X) of the liposomal system was obtained as Y = 236.5 x X

Recycling domains in plant cell morphogenesis: small GTPase effectors, plasma membrane signalling and the exocyst.

Science.gov (United States)

Zárský, Viktor; Potocký, Martin

2010-04-01

The Rho/Rop small GTPase regulatory module is central for initiating exocytotically ACDs (active cortical domains) in plant cell cortex, and a growing array of Rop regulators and effectors are being discovered in plants. Structural membrane phospholipids are important constituents of cells as well as signals, and phospholipid-modifying enzymes are well known effectors of small GTPases. We have shown that PLDs (phospholipases D) and their product, PA (phosphatidic acid), belong to the regulators of the secretory pathway in plants. We have also shown that specific NOXs (NADPH oxidases) producing ROS (reactive oxygen species) are involved in cell growth as exemplified by pollen tubes and root hairs. Most plant cells exhibit several distinct plasma membrane domains (ACDs), established and maintained by endocytosis/exocytosis-driven membrane protein recycling. We proposed recently the concept of a 'recycling domain' (RD), uniting the ACD and the connected endosomal recycling compartment (endosome), as a dynamic spatiotemporal entity. We have described a putative GTPase-effector complex exocyst involved in exocytic vesicle tethering in plants. Owing to the multiplicity of its Exo70 subunits, this complex, along with many RabA GTPases (putative recycling endosome organizers), may belong to core regulators of RD organization in plants.

MICOS and phospholipid transfer by Ups2-Mdm35 organize membrane lipid synthesis in mitochondria.

Science.gov (United States)

Aaltonen, Mari J; Friedman, Jonathan R; Osman, Christof; Salin, Bénédicte; di Rago, Jean-Paul; Nunnari, Jodi; Langer, Thomas; Tatsuta, Takashi

2016-06-06

Mitochondria exert critical functions in cellular lipid metabolism and promote the synthesis of major constituents of cellular membranes, such as phosphatidylethanolamine (PE) and phosphatidylcholine. Here, we demonstrate that the phosphatidylserine decarboxylase Psd1, located in the inner mitochondrial membrane, promotes mitochondrial PE synthesis via two pathways. First, Ups2-Mdm35 complexes (SLMO2-TRIAP1 in humans) serve as phosphatidylserine (PS)-specific lipid transfer proteins in the mitochondrial intermembrane space, allowing formation of PE by Psd1 in the inner membrane. Second, Psd1 decarboxylates PS in the outer membrane in trans, independently of PS transfer by Ups2-Mdm35. This latter pathway requires close apposition between both mitochondrial membranes and the mitochondrial contact site and cristae organizing system (MICOS). In MICOS-deficient cells, limiting PS transfer by Ups2-Mdm35 and reducing mitochondrial PE accumulation preserves mitochondrial respiration and cristae formation. These results link mitochondrial PE metabolism to MICOS, combining functions in protein and lipid homeostasis to preserve mitochondrial structure and function. © 2016 Aaltonen et al.

Modification of radiation-induced oxidative damage in liposomal and microsomal membrane by eugenol

Energy Technology Data Exchange (ETDEWEB)

Pandey, B.N. [Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Mumbai 400 085 (India); Lathika, K.M. [Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Mumbai 400 085 (India); Mishra, K.P. [Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Mumbai 400 085 (India)]. E-mail: kpm@magnum.barc.ernet.in

2006-03-15

Radiation-induced membrane oxidative damage, and their modification by eugenol, a natural antioxidant, was investigated in liposomes and microsomes. Liposomes prepared with DPH showed decrease in fluorescence after {gamma}-irradiation, which was prevented significantly by eugenol and correlated with magnitude of oxidation of phospholipids. Presence of eugenol resulted in substantial inhibition in MDA formation in irradiated liposomes/microsomes, which was less effective when added after irradiation. Similarly, the increase in phospholipase C activity observed after irradiation in microsomes was inhibited in samples pre-treated with eugenol. Results suggest association of radio- oxidative membrane damage with alterations in signaling molecules, and eugenol significantly prevented these membrane damaging events.

Characterising antimicrobial protein-membrane complexes

International Nuclear Information System (INIS)

Xun, Gloria; Dingley, Andrew; Tremouilhac, Pierre

2009-01-01

Full text: Antimicrobial proteins (AMPs) are host defence molecules that protect organisms from microbial infection. A number of hypotheses for AMP activity have been proposed which involve protein membrane interactions. However, there is a paucity of information describing AMP-membrane complexes in detail. The aim of this project is to characterise the interactions of amoebapore-A (APA-1) with membrane models using primarily solution-state NMR spectroscopy. APA-1 is an AMP which is regulated by a pH-dependent dimerisation event. Based on the atomic resolution solution structure of monomeric APA-1, it is proposed that this dimerisation is a prerequisite for ring-like hexameric pore formation. Due to the cytotoxicity of APA-1, we have developed a cell-free system to produce this protein. To facilitate our studies, we have adapted the cell-free system to isotope label APA-1. 13 C /15 N -enriched APA-1 sample was achieved and we have begun characterising APA-1 dimerisation and membrane interactions using NMR spectroscopy and other biochemical/biophysical methods. Neutron reflectometry is a surface-sensitive technique and therefore represents an ideal technique to probe how APA-1 interacts with membranes at the molecular level under different physiological conditions. Using Platypus, the pH-induced APA-1-membrane interactions should be detectable as an increase of the amount of protein adsorbed at the membrane surface and changes in the membrane properties. Specifically, detailed information of the structure and dimensions of the protein-membrane complex, the position and amount of the protein in the membrane, and the perturbation of the membrane phospholipids on protein incorporation can be extracted from the neutron reflectometry measurement. Such information will enable critical assessment of current proposed mechanisms of AMP activity in bacterial membranes and complement our NMR studies

High Fat Diet-Induced Changes in Mouse Muscle Mitochondrial Phospholipids Do Not Impair Mitochondrial Respiration Despite Insulin Resistance

Science.gov (United States)

Hulshof, Martijn F. M.; van den Berg, Sjoerd A. A.; Schaart, Gert; van Dijk, Ko Willems; Smit, Egbert; Mariman, Edwin C. M.

2011-01-01

Background Type 2 diabetes mellitus and muscle insulin resistance have been associated with reduced capacity of skeletal muscle mitochondria, possibly as a result of increased intake of dietary fat. Here, we examined the hypothesis that a prolonged high-fat diet consumption (HFD) increases the saturation of muscle mitochondrial membrane phospholipids causing impaired mitochondrial oxidative capacity and possibly insulin resistance. Methodology C57BL/6J mice were fed an 8-week or 20-week low fat diet (10 kcal%; LFD) or HFD (45 kcal%). Skeletal muscle mitochondria were isolated and fatty acid (FA) composition of skeletal muscle mitochondrial phospholipids was analyzed by thin-layer chromatography followed by GC. High-resolution respirometry was used to assess oxidation of pyruvate and fatty acids by mitochondria. Insulin sensitivity was estimated by HOMA-IR. Principal Findings At 8 weeks, mono-unsaturated FA (16∶1n7, 18∶1n7 and 18∶1n9) were decreased (−4.0%, p<0.001), whereas saturated FA (16∶0) were increased (+3.2%, p<0.001) in phospholipids of HFD vs. LFD mitochondria. Interestingly, 20 weeks of HFD descreased mono-unsaturated FA while n-6 poly-unsaturated FA (18∶2n6, 20∶4n6, 22∶5n6) showed a pronounced increase (+4.0%, p<0.001). Despite increased saturation of muscle mitochondrial phospholipids after the 8-week HFD, mitochondrial oxidation of both pyruvate and fatty acids were similar between LFD and HFD mice. After 20 weeks of HFD, the increase in n-6 poly-unsaturated FA was accompanied by enhanced maximal capacity of the electron transport chain (+49%, p = 0.002) and a tendency for increased ADP-stimulated respiration, but only when fuelled by a lipid-derived substrate. Insulin sensitivity in HFD mice was reduced at both 8 and 20 weeks. Conclusions/Interpretation Our findings do not support the concept that prolonged HF feeding leads to increased saturation of skeletal muscle mitochondrial phospholipids resulting in a decrease in

High fat diet-induced changes in mouse muscle mitochondrial phospholipids do not impair mitochondrial respiration despite insulin resistance.

Directory of Open Access Journals (Sweden)

Joris Hoeks

Full Text Available BACKGROUND: Type 2 diabetes mellitus and muscle insulin resistance have been associated with reduced capacity of skeletal muscle mitochondria, possibly as a result of increased intake of dietary fat. Here, we examined the hypothesis that a prolonged high-fat diet consumption (HFD increases the saturation of muscle mitochondrial membrane phospholipids causing impaired mitochondrial oxidative capacity and possibly insulin resistance. METHODOLOGY: C57BL/6J mice were fed an 8-week or 20-week low fat diet (10 kcal%; LFD or HFD (45 kcal%. Skeletal muscle mitochondria were isolated and fatty acid (FA composition of skeletal muscle mitochondrial phospholipids was analyzed by thin-layer chromatography followed by GC. High-resolution respirometry was used to assess oxidation of pyruvate and fatty acids by mitochondria. Insulin sensitivity was estimated by HOMA-IR. PRINCIPAL FINDINGS: At 8 weeks, mono-unsaturated FA (16∶1n7, 18∶1n7 and 18∶1n9 were decreased (-4.0%, p<0.001, whereas saturated FA (16∶0 were increased (+3.2%, p<0.001 in phospholipids of HFD vs. LFD mitochondria. Interestingly, 20 weeks of HFD descreased mono-unsaturated FA while n-6 poly-unsaturated FA (18∶2n6, 20∶4n6, 22∶5n6 showed a pronounced increase (+4.0%, p<0.001. Despite increased saturation of muscle mitochondrial phospholipids after the 8-week HFD, mitochondrial oxidation of both pyruvate and fatty acids were similar between LFD and HFD mice. After 20 weeks of HFD, the increase in n-6 poly-unsaturated FA was accompanied by enhanced maximal capacity of the electron transport chain (+49%, p = 0.002 and a tendency for increased ADP-stimulated respiration, but only when fuelled by a lipid-derived substrate. Insulin sensitivity in HFD mice was reduced at both 8 and 20 weeks. CONCLUSIONS/INTERPRETATION: Our findings do not support the concept that prolonged HF feeding leads to increased saturation of skeletal muscle mitochondrial phospholipids resulting in a decrease in

The lipid organisation of the cell membrane

Directory of Open Access Journals (Sweden)

Ladha, S.

2000-04-01

Full Text Available Lipids and proteins in biological membranes are arranged in a mosaic of domains in the membrane. These domains represent small-scale heterogeneities in composition, shape and fluidity within the plane of the membrane, over the range of hundreds of nanometers to a few micrometers. They arise from the complex interactions of the heterogeneous mixtures of phospholipids, sterols, and proteins that make up all biological membranes.Los lípidos y las proteínas en las membranas biológicas están dispuestos en un mosaico de campos en la membrana. Estos campos representan heterogeneidades a pequeña escala en la composición, forma y fluidez dentro del plano de la membrana, en un rango que va de los cientos de nanómetros a los pocos micrómetros. Estos campos se originan de las complejas interacciones de las mezclas heterogéneas de fosfolípidos, esteroles y proteínas de las que están hechas todas y cada una de las membranas biológicas.

Composition and metabolism of phospholipids in Octopus vulgaris and Sepia officinalis hatchlings.

Science.gov (United States)

Reis, Diana B; Acosta, Nieves G; Almansa, Eduardo; Tocher, Douglas R; Andrade, José P; Sykes, António V; Rodríguez, Covadonga

2016-10-01

The objective of the present study was to characterise the fatty acid (FA) profiles of the major phospholipids, of Octopus vulgaris and Sepia officinalis hatchlings, namely phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylinositol (PI) and phosphatidylethanolamine (PE); and to evaluate the capability of both cephalopod species on dietary phospholipid remodelling. Thus, O. vulgaris and S. officinalis hatchlings were in vivo incubated with 0.3μM of L-∝-1-palmitoyl-2-[1-(14)C]arachidonyl-PC or L-∝-1-palmitoyl-2-[1-(14)C]arachidonyl-PE. Octopus and cuttlefish hatchlings phospholipids showed a characteristic FA profiles with PC presenting high contents of 16:0 and 22:6n-3 (DHA); PS having high 18:0, DHA and 20:5n-3 (EPA); PI a high content of saturated FA; and PE showing high contents of DHA and EPA. Interestingly, the highest content of 20:4n-6 (ARA) was found in PE rather than PI. Irrespective of the phospholipid in which [1-(14)C]ARA was initially bound (either PC or PE), the esterification pattern of [1-(14)C]ARA in octopus lipids was similar to that found in their tissues with high esterification of this FA into PE. In contrast, in cuttlefish hatchlings [1-(14)C]ARA was mainly recovered in the same phospholipid that was provided. These results showed a characteristic FA profiles in the major phospholipids of the two species, as well as a contrasting capability to remodel dietary phospholipids, which may suggest a difference in phospholipase activities. Copyright © 2016 Elsevier Inc. All rights reserved.

Exit-strategies - smart ways to release phospholipid vesicle cargo

OpenAIRE

Mellal Denia; Zumbuehl Andreas

2014-01-01

This highlight describes recent trends in fundamental phospholipid research towards possible future drug delivery technology. In particular it focuses on synthetic phospholipids and their vesicular constructs and describes selected “smart” ways to release cargo from liposomes. Various chemical and physical release triggers are discussed such as temperature changes, application of ultrasound, enzyme degradation, changes in pH, redox reactions, photochemical reactions, as well as the effects of...

Binding of the GTPase Sar1 to a Lipid Membrane Monolayer: Insertion and Orientation Studied by Infrared Reflection–Absorption Spectroscopy

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Christian Schwieger

2017-11-01

Full Text Available Membrane-interacting proteins are polyphilic polymers that engage in dynamic protein–protein and protein–lipid interactions while undergoing changes in conformation, orientation and binding interfaces. Predicting the sites of interactions between such polypeptides and phospholipid membranes is still a challenge. One example is the small eukaryotic GTPase Sar1, which functions in phospholipid bilayer remodeling and vesicle formation as part of the multimeric coat protein complex (COPII. The membrane interaction of Sar1 is strongly dependent on its N-terminal 23 amino acids. By monolayer adsorption experiments and infrared reflection-absorption spectroscopy (IRRAS, we elucidate the role of lipids in inducing the amphipathicity of this N-terminal stretch, which inserts into the monolayer as an amphipathic helix (AH. The AH inserting angle is determined and is consistent with the philicities and spatial distribution of the amino acid monomers. Using an advanced method of IRRAS data evaluation, the orientation of Sar1 with respect to the lipid layer prior to the recruitment of further COPII proteins is determined. The result indicates that only a slight reorientation of the membrane-bound Sar1 is needed to allow coat assembly. The time-course of the IRRAS analysis corroborates a role of slow GTP hydrolysis in Sar1 desorption from the membrane.

Soft contact lens biomaterials from bioinspired phospholipid polymers.

Science.gov (United States)

Goda, Tatsuro; Ishihara, Kazuhiko

2006-03-01

Soft contact lens (SCL) biomaterials originated from the discovery of a poly(2-hydroxyethyl methacrylate) (poly[HEMA])-based hydrogel in 1960. Incorporation of hydrophilic polymers into poly(HEMA) hydrogels was performed in the 1970-1980s, which brought an increase in the equilibrium water content, leading to an enhancement of the oxygen permeability. Nowadays, the poly(HEMA)-based hydrogels have been applied in disposable SCL. At the same time, high oxygen-permeable silicone hydrogels were produced, which made it possible to continually wear SCL. Recently, numerous trials for improving the water wettability of silicone hydrogels have been performed. However, little attention has been paid to improving their anti-biofouling properties and biocompatibility. Since biomimetic phospholipid polymers possess excellent anti-biofouling properties and biocompatibility they have the potential to play a valuable role in the surface modification of the silicone hydrogel. The representative phospholipid polymers containing a 2-methacryloyloxyethyl phosphorylcholine (MPC) unit suppressed nonspecific protein adsorption, increased cell compatibility and contributed to blood compatible biomaterials. The MPC polymer coating on the silicone hydrogel improved its water wettability and biocompatibility, while maintaining high oxygen permeability compared with the original silicone hydrogel. Furthermore, the newly prepared phospholipid-type intermolecular crosslinker made it possible to synthesize a 100% phospholipid polymer hydrogel that can enhance the anti-biofouling properties and biocompatibility. In this review, the authors discuss how polymer hydrogels should be designed in order to obtain a biocompatible SCL and future perspectives.

Complete dissociation of the HIV-1 gp41 ectodomain and membrane proximal regions upon phospholipid binding

International Nuclear Information System (INIS)

Roche, Julien; Louis, John M.; Aniana, Annie; Ghirlando, Rodolfo; Bax, Ad

2015-01-01

The envelope glycoprotein gp41 mediates the process of membrane fusion that enables entry of the HIV-1 virus into the host cell. Strong lipid affinity of the ectodomain suggests that its heptad repeat regions play an active role in destabilizing membranes by directly binding to the lipid bilayers and thereby lowering the free-energy barrier for membrane fusion. In such a model, immediately following the shedding of gp120, the N-heptad and C-heptad helices dissociate and melt into the host cell and viral membranes, respectively, pulling the destabilized membranes into juxtaposition, ready for fusion. Post-fusion, reaching the final 6-helix bundle (6HB) conformation then involves competition between intermolecular interactions needed for formation of the symmetric 6HB trimer and the membrane affinity of gp41’s ectodomain, including its membrane-proximal regions. Our solution NMR study of the structural and dynamic properties of three constructs containing the ectodomain of gp41 with and without its membrane-proximal regions suggests that these segments do not form inter-helical interactions until the very late steps of the fusion process. Interactions between the polar termini of the heptad regions, which are not associating with the lipid surface, therefore may constitute the main driving force initiating formation of the final post-fusion states. The absence of significant intermolecular ectodomain interactions in the presence of dodecyl phosphocholine highlights the importance of trimerization of gp41’s transmembrane helix to prevent complete dissociation of the trimer during the course of fusion

Complete dissociation of the HIV-1 gp41 ectodomain and membrane proximal regions upon phospholipid binding

Energy Technology Data Exchange (ETDEWEB)

Roche, Julien; Louis, John M.; Aniana, Annie [National Institute of Diabetes and Digestive and Kidney Diseases, Laboratory of Chemical Physics (United States); Ghirlando, Rodolfo [National Institutes of Health, Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases (United States); Bax, Ad, E-mail: bax@nih.gov [National Institute of Diabetes and Digestive and Kidney Diseases, Laboratory of Chemical Physics (United States)

2015-04-15

The envelope glycoprotein gp41 mediates the process of membrane fusion that enables entry of the HIV-1 virus into the host cell. Strong lipid affinity of the ectodomain suggests that its heptad repeat regions play an active role in destabilizing membranes by directly binding to the lipid bilayers and thereby lowering the free-energy barrier for membrane fusion. In such a model, immediately following the shedding of gp120, the N-heptad and C-heptad helices dissociate and melt into the host cell and viral membranes, respectively, pulling the destabilized membranes into juxtaposition, ready for fusion. Post-fusion, reaching the final 6-helix bundle (6HB) conformation then involves competition between intermolecular interactions needed for formation of the symmetric 6HB trimer and the membrane affinity of gp41’s ectodomain, including its membrane-proximal regions. Our solution NMR study of the structural and dynamic properties of three constructs containing the ectodomain of gp41 with and without its membrane-proximal regions suggests that these segments do not form inter-helical interactions until the very late steps of the fusion process. Interactions between the polar termini of the heptad regions, which are not associating with the lipid surface, therefore may constitute the main driving force initiating formation of the final post-fusion states. The absence of significant intermolecular ectodomain interactions in the presence of dodecyl phosphocholine highlights the importance of trimerization of gp41’s transmembrane helix to prevent complete dissociation of the trimer during the course of fusion.

Milk phospholipid's protective effects against UV damage in skin equivalent models

Science.gov (United States)

Dargitz, Carl; Russell, Ashley; Bingham, Michael; Achay, Zyra; Jimenez-Flores, Rafael; Laiho, Lily H.

2012-03-01

Exposure of skin tissue to UV radiation has been shown to cause DNA photodamage. If this damaged DNA is allowed to replicate, carcinogenesis may occur. DNA damage is prevented from being passed on to daughter cells by upregulation of the protein p21. p21 halts the cells cycle allowing the cell to undergo apoptosis, or repair its DNA before replication. Previous work suggested that milk phospholipids may possess protective properties against UV damage. In this study, we observed cell morphology, cell apoptosis, and p21 expression in tissue engineered epidermis through the use of Hematoxylin and Eosin staining, confocal microscopy, and western blot respectively. Tissues were divided into four treatment groups including: a control group with no UV and no milk phospholipid treatment, a group exposed to UV alone, a group incubated with milk phospholipids alone, and a group treated with milk phospholipids and UV. All groups were incubated for twenty-four hours after treatment. Tissues were then fixed, processed, and embedded in paraffin. Performing western blots resulted in visible p21 bands for the UV group only, implying that in every other group, p21 expression was lesser. Numbers of apoptotic cells were determined by observing the tissues treated with Hoechst dye under a confocal microscope, and counting the number of apoptotic and total cells to obtain a percentage of apoptotic cells. We found a decrease in apoptotic cells in tissues treated with milk phospholipids and UV compared to tissues exposed to UV alone. Collectively, these results suggest that milk phospholipids protect cell DNA from damage incurred from UV light.

Beneficial effects of gamma linolenic acid supplementation on nerve conduction velocity, Na+, K+ ATPase activity, and membrane fatty acid composition in sciatic nerve of diabetic rats.

Science.gov (United States)

Coste, T; Pierlovisi, M; Leonardi, J; Dufayet, D; Gerbi, A; Lafont, H; Vague, P; Raccah, D

1999-07-01

Metabolic and vascular abnormalities are implicated in the pathogenesis of diabetic neuropathy. Two principal metabolic defects are altered lipid metabolism resulting from the impairment of delta-6-desaturase, which converts linoleic acid (LA) into gamma linolenic acid (GLA), and reduced nerve Na+, K+ ATPase activity. This reduction may be caused by a lack of incorporation of (n-6) fatty acids in membrane phospholipids. Because this ubiquitous enzyme maintains the membrane electrical potential and allows repolarization, disturbances in its activity can alter the process of nerve conduction velocity (NCV). We studied the effects of supplementation with GLA (260 mg per day) on NCV, fatty acid phospholipid composition, and Na+, K+ ATPase activity in streptozotocin-diabetic rats. Six groups of 10 rats were studied. Two groups served as controls supplemented with GLA or sunflower oil (GLA free). Two groups with different durations of diabetes were studied: 6 weeks with no supplementation and 12 weeks supplemented with sunflower oil. To test the ability of GLA to prevent or reverse the effects of diabetes, two groups of diabetic rats were supplemented with GLA, one group for 12 weeks and one group for 6 weeks, starting 6 weeks after diabetes induction. Diabetes resulted in a 25% decrease in NCV (P < 0.0001), a 45% decrease in Na+, K+ ATPase activity (P < 0.0001), and an abnormal phospholipid fatty acid composition. GLA restored NCV both in the prevention and reversal studies and partially restored Na+, K+ ATPase activity in the preventive treatment group (P < 0.0001). These effects were accompanied by a modification of phospholipid fatty acid composition in nerve membranes. Overall, the results suggest that membrane fatty acid composition plays a direct role in NCV and confirm the beneficial effect of GLA supplementation in diabetic neuropathy.

Proton conductance at elevated temperature:Formulation and investigation of poly(4-styrenesulfonic acid / 4-aminobenzylamine / phosphoric acid membranes

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Jalal eJalili

2014-07-01

Full Text Available 4-aminobenzylamine and phosphoric acid were blended in various proportions with poly (4-styrenesulfonic acid to form a new group of membranes exhibiting proton conductance under water-free conditions. The 4-aminobenzylamine molecule, possessing an aniline-like and benzylamine-like functional group, can interact both with the phosphoric acid and the poly(4-styrenesulfonic acid via nucleophilic interaction, thereby allowing proton jumping in the structure. Physico-chemical and thermal characteristics of the prepared solid membranes were investigated by IR spectroscopy and thermo-gravimetric analysis, respectively. Electrochemical impedance spectroscopy was employed to investigate their proton-conductance properties. Transparent composite membranes were prepared. However, the membranes are opaque for relatively high content of phosphoric acid. These membranes are thermally stable up to 300°C. The proton conductivity increases with temperature and also with content of phosphoric acid. Values as high as 1.8×10–3 S cm–1 were measured at 190°C in fully anhydrous condition.

Precise detection of pH inside large unilamellar vesicles using membrane-impermeable dendritic porphyrin-based nanoprobes.

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Leiding, Thom; Górecki, Kamil; Kjellman, Tomas; Vinogradov, Sergei A; Hägerhäll, Cecilia; Arsköld, Sindra Peterson

2009-05-15

Accurate real-time measurements of proton concentration gradients are pivotal to mechanistic studies of proton translocation by membrane-bound enzymes. Here we report a detailed characterization of the pH-sensitive fluorescent nanoprobe Glu(3), which is well suited for pH measurements in microcompartmentalized biological systems. The probe is a polyglutamic porphyrin dendrimer in which multiple carboxylate termini ensure its high water solubility and prevent its diffusion across phospholipid membranes. The probe's pK is in the physiological pH range, and its protonation can be followed ratiometrically by absorbance or fluorescence in the ultraviolet-visible spectral region. The usefulness of the probe was enhanced by using a semiautomatic titration system coupled to a charge-coupled device (CCD) spectrometer, enabling fast and accurate titrations and full spectral coverage of the system at millisecond time resolution. The probe's pK was measured in bulk solutions as well as inside large unilamellar vesicles in the presence of physiologically relevant ions. Glu(3) was found to be completely membrane impermeable, and its distinct spectroscopic features permit pH measurements inside closed membrane vesicles, enabling quantitative mechanistic studies of membrane-spanning proteins. Performance of the probe was demonstrated by monitoring the rate of proton leakage through the phospholipid bilayer in large vesicles with and without the uncoupler gramicidin present. Overall, as a probe for biological proton translocation measurements, Glu(3) was found to be superior to the commercially available pH indicators.

Tissue phospholipids (TPL) in avian tuberculosis (AT)

International Nuclear Information System (INIS)

Nandedkar, A.K.N.; Malhotra, H.C.

1986-01-01

AT constitutes one of the major problems in animal husbandry. Chickens (white, leg horn, male, 400-600 g) were infected with Mycobacterium avium maintained on I.U.T. medium to induce clinical AT which was confirmed by histopathological examinations of the affected tissues. Fatty infiltration and tissue enlargement was visible in infected birds. After 4 wks, incorporation of i.v. 32 P (50 uCi/100 g body wt.) in affected tissues was followed for 3,7,9,12 hr intervals. Lipids were extracted and fractionated by silicic acid (SA) column and SA impregnated paper chromatography. When compared with pair-fed controls, in AT slower turnover of TPL in liver, slightly higher in heart and significantly increased turnover of TPL in serum were observed. No appreciable change in total TPL content was noticed in brain, spleen and kidney. Further fractionation of TPL provided better understanding of the metabolism. Increase in lysophosphatidyl-choline (LPC) and -ethanolamine (LPE) content, powerful hemolytic agents, in liver may explain frequent occurrence of anemia in tuberculosis. Also, a concomitant marked increase in the ratio of total saturated/unsaturated fatty acids is observed in serum phosphatidyl choline fraction. This confirms the observation that the membrane phospholipid metabolism is significantly affected in tuberculosis infection

Effect of free cholesterol on incorporation of triolein in phospholipid bilayers

International Nuclear Information System (INIS)

Spooner, P.J.R.; Small, D.M.

1987-01-01

Triacylglycerols are the major substrates for lipolytic enzymes that act at the surface of emulsion-like particles such as triglyceride-rich lipoproteins, chylomicrons, and intracellular lipid droplets. This study examines the effect of cholesterol on the solubility of a triacylglycerol, triolein, in phospholipid surfaces. Solubilities of [carbonyl- 13 C] triolein in phospholipid bilayer vesicles containing between 0 and 50 mol % free cholesterol, prepared by cosonication, were measured by 13 C NMR. The carbonyl resonances from bilayer-incorporated triglyceride were shifted downfield in the 13 C NMR spectra from those corresponding to excess, nonincorporated material. This enabled solubilities to be determined directly from carbonyl peak intensities at most cholesterol concentration. The bilayer solubility of triolein was inversely proportional to the cholesterol/phospholipid mole ratio. In pure phospholipid vesicles the triolein solubility was 2.2 mol %. The triglyceride incorporation decreased to 1.1 mol % at a cholesterol/phospholipid mole ratio of 0.5, and at a mole ratio of 1.0 for the bilayer lipids, the triolein solubility was reduced to just 0.15 mol %. The effects of free cholesterol were more pronounced and progressive than observed previously on the bilayer solubility of cholestery oleate. As with cholesteryl oleate, they suggest that cholesterol also displaces solubilized triglyceride to deeper regions of the bilayer

Critical roles of isoleucine-364 and adjacent residues in a hydrophobic gate control of phospholipid transport by the mammalian P4-ATPase ATP8A2.

Science.gov (United States)

Vestergaard, Anna L; Coleman, Jonathan A; Lemmin, Thomas; Mikkelsen, Stine A; Molday, Laurie L; Vilsen, Bente; Molday, Robert S; Dal Peraro, Matteo; Andersen, Jens Peter

2014-04-08

P4-ATPases (flippases) translocate specific phospholipids such as phosphatidylserine from the exoplasmic leaflet of the cell membrane to the cytosolic leaflet, upholding an essential membrane asymmetry. The mechanism of flipping this giant substrate has remained an enigma. We have investigated the importance of amino acid residues in transmembrane segment M4 of mammalian P4-ATPase ATP8A2 by mutagenesis. In the related ion pumps Na(+),K(+)-ATPase and Ca(2+)-ATPase, M4 moves during the enzyme cycle, carrying along the ion bound to a glutamate. In ATP8A2, the corresponding residue is an isoleucine, which recently was found mutated in patients with cerebellar ataxia, mental retardation, and dysequilibrium syndrome. Our analyses of the lipid substrate concentration dependence of the overall and partial reactions of the enzyme cycle in mutants indicate that, during the transport across the membrane, the phosphatidylserine head group passes near isoleucine-364 (I364) and that I364 is critical to the release of the transported lipid into the cytosolic leaflet. Another M4 residue, N359, is involved in recognition of the lipid substrate on the exoplasmic side. Our functional studies are supported by structural homology modeling and molecular dynamics simulations, suggesting that I364 and adjacent hydrophobic residues function as a hydrophobic gate that separates the entry and exit sites of the lipid and directs sequential formation and annihilation of water-filled cavities, thereby enabling transport of the hydrophilic phospholipid head group in a groove outlined by the transmembrane segments M1, M2, M4, and M6, with the hydrocarbon chains following passively, still in the membrane lipid phase.

Determination of phospholipid regiochemistry by Ag(I) adduction and tandem mass spectrometry.

Science.gov (United States)

Yoo, Hyun Ju; Håkansson, Kristina

2011-02-15

Collision-activated dissociation (CAD) and infrared multiphoton dissociation (IRMPD) of Ag-adducted phospholipids were investigated as structural tools. Previously, determination of the acyl chains at the two phospholipid esterification sites has been performed based on the R(1)COO(-)/R(2)COO(-) ratio in negative ion mode CAD tandem mass spectrometry. However, the observed product ion ratio is dependent on the extent of unsaturation of the fatty acyl group at sn-2 as well as on the total chain length. Similarly, in positive ion mode CAD with/without alkaline or alkaline earth metal adduction, the ratio of product ions resulting from either R(1)COOH or R(2)COOH neutral losses is dependent on the nature of the phospholipid polar headgroup. Ag(+) ion chromatography, in which silver ions are part of the stationary phase, can provide information on double bond number/distribution as well as double bond configuration (cis/trans) because of interaction between Ag(+) ions and olefinic π electrons of fatty acids and lipids. We hypothesized that interactions between double bonds and Ag(+) may be utilized to also reveal phospholipid esterification site information in tandem mass spectrometry. CAD and IRMPD of Ag-adducted phospholipids with unsaturated fatty acids (R(x)COOH, x = 1 or 2) provided characteristic product ions, [R(x)COOH + Ag](+), and their neutral losses. The characteristic product ions and their abundances do not depend on the type of polar headgroup or the number of double bonds of unsaturated acyl chains. Tandem mass spectrometry of Cu-adducted phospholipids was also performed for comparison based on the Lewis acid and base properties of Cu(+) and phospholipid double bonds, respectively.

Direct investigation of the vectorization properties of amphiphilic cyclodextrins in phospholipid films.

Science.gov (United States)

Javierre, Isabelle; Nedyalkov, Mickael; Petkova, Vera; Benattar, Jean Jacques; Weisse, Sandrine; Auzély-Velty, Rachel; Djedaïni-Pilard, Florence; Perly, Bruno

2002-10-01

Recently, new cyclodextrin derivatives were synthesized and shown to exhibit strong amphiphilic properties. In this paper, we study the action of these new amphiphilic cyclodextrins on phospholipids. Mixed phospholipid/cyclodextrin derivative films were prepared and studied using X-ray reflectivity for various phospholipid/cyclodextrin ratios. A molar ratio of 3 provides a highly stable film the molecular structure of which has been investigated in detail. The cholesterol tail of the cyclodextrin molecule was found to be anchored into the phospholipid film. The cyclodextrin moieties exposed to the aqueous medium are prone to the addition of the guest molecule Dosulepin, making them of high interest for drug delivery. For this purpose and as an example of a potential application, this cyclodextrin molecular carrier property is also addressed to this complex film architecture.

A retrospective: Use of Escherichia coli as a vehicle to study phospholipid synthesis and function

Science.gov (United States)

Dowhan, William

2012-01-01

Although the study of individual phospholipids and their synthesis began in the 1920’s first in plants and then mammals, it was not until the early 1960’s that Eugene Kennedy using Escherichia coli initiated studies of bacterial phospholipid metabolism. With the base of information already available from studies of mammalian tissue, the basic blueprint of phospholipid biosynthesis in E. coli was worked out by the late 1960’s. In 1970’s and 1980’s most of the enzymes responsible for phospholipid biosynthesis were purified and many of the genes encoding these enzymes were identified. By the late 1990’s conditional and null mutants were available along with clones of the genes for every step of phospholipid biosynthesis. Most of these genes had been sequenced before the complete E. coli genome sequence was available. Strains of E. coli were developed in which phospholipid composition could be changed in a systematic manner while maintaining cell viability. Null mutants, strains in which phospholipid metabolism was artificially regulated, and strains synthesizing foreign lipids not found in E. coli have been used to this day to define specific roles for individual phospholipid. This review will trace the findings that have led to the development of E. coli as an excellent model system to study mechanisms underlying the synthesis and function of phospholipids that are widely applicable to other prokaryotic and eukaryotic systems. PMID:22925633

Insertion of Neurotransmitters into a Lipid Bilayer Membrane and Its Implication on Membrane Stability: A Molecular Dynamics Study.

Science.gov (United States)

Shen, Chun; Xue, Minmin; Qiu, Hu; Guo, Wanlin

2017-03-17

The signaling molecules in neurons, called neurotransmitters, play an essential role in the transportation of neural signals, during which the neurotransmitters interact with not only specific receptors, but also cytomembranes, such as synaptic vesicle membranes and postsynaptic membranes. Through extensive molecular dynamics simulations, the atomic-scale insertion dynamics of typical neurotransmitters, including methionine enkephalin (ME), leucine enkephalin (LE), dopamine (DA), acetylcholine (ACh), and aspartic acid (ASP), into lipid bilayers is investigated. The results show that the first three neurotransmitters (ME, LE, and DA) are able to diffuse freely into both 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) membranes, and are guided by the aromatic residues Tyr and Phe. Only a limited number of these neurotransmitters are allowed to penetrate into the membrane, which suggests an intrinsic mechanism by which the membrane is protected from being destroyed by excessive inserted neurotransmitters. After spontaneous insertion, the neurotransmitters disturb the surrounding phospholipids in the membrane, as indicated by the altered distribution of components in lipid leaflets and the disordered lipid tails. In contrast, the last two neurotransmitters (ACh and ASP) cannot enter the membrane, but instead always diffuse freely in solution. These findings provide an understanding at the atomic level of how neurotransmitters interact with the surrounding cytomembrane, as well as their impact on membrane behavior. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Self-assembly of nanoscale particles with biosurfactants and membrane scaffold proteins.

Science.gov (United States)

Faas, Ramona; Pohle, Annelie; Moß, Karin; Henkel, Marius; Hausmann, Rudolf

2017-12-01

Nanodiscs are membrane mimetics which may be used as tools for biochemical and biophysical studies of a variety of membrane proteins. These nanoscale structures are composed of a phospholipid bilayer held together by an amphipathic membrane scaffold protein (MSP). In the past, nanodiscs were successfully assembled with membrane scaffold protein 1D1 and 1,2-dipalmitoyl- sn -glycero-3-phosphorylcholine with a homogeneous diameter of ∼10 nm. In this study, the formation of nanoscale particles from MSP1D1 and rhamnolipid biosurfactants is investigated. Different protein to lipid ratios of 1:80, 1:90 and 1:100 were used for the assembly reaction, which were consecutively separated, purified and analyzed by size-exclusion chromatography (SEC) and dynamic light scattering (DLS). Size distributions were measured to determine homogeneity and confirm size dimensions. In this study, first evidence is presented on the formation of nanoscale particles with rhamnolipid biosurfactants and membrane scaffold proteins.

Estimation of the Asymmetrical Arrangement of Plasma Membrane Aminophospholipids. An Experimental Assay for Students of Biochemistry and Molecular Biology.

Science.gov (United States)

Sanchez-Yague, J.; And Others

1987-01-01

Describes an experiment to discover the topology of plasma membrane aminophospholipids (phosphatidylethanolamine and phosphatidylserine) using whole platelets and trinitrobenzene sulfonate (TNBS) as a probe. Reports changes in phospholipid distribution during platelet activation with simultaneous action of thrombia and collagen. Details the…

The Inner Membrane Complex Sub-compartment Proteins Critical for Replication of the Apicomplexan Parasite Toxoplasma gondii Adopt a Pleckstrin Homology Fold*

Science.gov (United States)

Tonkin, Michelle L.; Beck, Josh R.; Bradley, Peter J.; Boulanger, Martin J.

2014-01-01

Toxoplasma gondii, an apicomplexan parasite prevalent in developed nations, infects up to one-third of the human population. The success of this parasite depends on several unique structures including an inner membrane complex (IMC) that lines the interior of the plasma membrane and contains proteins important for gliding motility and replication. Of these proteins, the IMC sub-compartment proteins (ISPs) have recently been shown to play a role in asexual T. gondii daughter cell formation, yet the mechanism is unknown. Complicating mechanistic characterization of the ISPs is a lack of sequence identity with proteins of known structure or function. In support of elucidating the function of ISPs, we first determined the crystal structures of representative members TgISP1 and TgISP3 to a resolution of 2.10 and 2.32 Å, respectively. Structural analysis revealed that both ISPs adopt a pleckstrin homology fold often associated with phospholipid binding or protein-protein interactions. Substitution of basic for hydrophobic residues in the region that overlays with phospholipid binding in related pleckstrin homology domains, however, suggests that ISPs do not retain phospholipid binding activity. Consistent with this observation, biochemical assays revealed no phospholipid binding activity. Interestingly, mapping of conserved surface residues combined with crystal packing analysis indicates that TgISPs have functionally repurposed the phospholipid-binding site likely to coordinate protein partners. Recruitment of larger protein complexes may also be aided through avidity-enhanced interactions resulting from multimerization of the ISPs. Overall, we propose a model where TgISPs recruit protein partners to the IMC to ensure correct progression of daughter cell formation. PMID:24675080

Phospholipid analogue distributions of Iranian isolates of candida

International Nuclear Information System (INIS)

Zarei Mahmoudabadi, A.; Brucker, D.B.

2004-01-01

The aim of this study was to analyse polar lipids of candida species isolated from Ahwas (Iran) by fast Atom bombardment mass spectrometry . Nine isolates of Candida Sp. were identified by growth at 45 d ig c , production of chlamydoconidia on cornmeal agar, colonial colour on CHROMagar Candida, germ tube production and ID 32 C kits. Then polar lipids were extracted from freeze-dried cultures and analysed using Fast Atom Bombardment Mass Spectrometry. The most intense carboxylate and phospholipid molecular species anions were of m/z 281 (C 1 8 : 1 ) and m/z 515 (PA 23:2). However, the most intense carboxylate and phospholipid analogues in Candida Parapsilosis were 292 (Un) and 555 (PA 26:3), which differed from other yeasts. Isolates were grouped by single linkage clustering based on correlation coefficient for strain pairs calculated with carboxylate and phospholipid molecular species distributions. Fast Atom Bombardment Mass Spectrometry can differentiate the C. albicans based on analysis of polar lipid distributions.These findings support that differentiation between C. albicans and other species is possible based on polar lipids

Effects of ionizing radiation on the peroxide content of a pure polyunsaturated lipid dispersion and of lipids and membranes derived from Acholeplasma laidlawii

Energy Technology Data Exchange (ETDEWEB)

Edwards, J.C.; Cramp, W.A. (Hammersmith Hospital, London (UK). M.R.C. Cyclotron Unit); Chapman, D. (Royal Free Hospital, London (UK))

1984-01-01

Dispersions of a pure unsaturated phospholipid, dilinoleoylphosphatidyl choline, formed conjugated diene hydroperoxides when irradiated in air with 7 MeV electrons (150 Gy and 300 Gy). Peroxide formation was optimized when the dispersions were irradiated in air at 37/sup 0/C at a dose rate of 5 Gy/min. No significant loss of linoleic acid from the irradiated phospholipid dispersions was observed after doses of 150 or 300 Gy. Small amounts of thiobarbituric acid-reactive material were formed in irradiated unsaturated phospholipid dispersions. However, lipids or membranes isolated from 48 hour cultures of Acholeplasma laidlawii grown in media supplemented with either linoleic or linolenic acid did not appear to be peroxidized by irradiation under the same conditions.

Sterol transfer between cyclodextrin and membranes: similar but not identical mechanism to NPC2-mediated cholesterol transfer.

Science.gov (United States)

McCauliff, Leslie A; Xu, Zhi; Storch, Judith

2011-08-30

Niemann--Pick C disease is an inherited disorder in which cholesterol and other lipids accumulate in the late endosomal/lysosomal compartment. Recently, cyclodextrins (CD) have been shown to reduce symptoms and extend lifespan in animal models of the disease. In the present studies we examined the mechanism of sterol transport by CD using in vitro model systems and fluorescence spectroscopy and NPC2-deficient fibroblasts. We demonstrate that cholesterol transport from the lysosomal cholesterol-binding protein NPC2 to CD occurs via aqueous diffusional transfer and is very slow; the rate-limiting step appears to be dissociation of cholesterol from NPC2, suggesting that specific interactions between NPC2 and CD do not occur. In contrast, the transfer rate of the fluorescent cholesterol analogue dehydroergosterol (DHE) from CD to phospholipid membranes is very rapid and is directly proportional to the acceptor membrane concentration, as is DHE transfer from membranes to CD. Moreover, CD dramatically increases the rate of sterol transfer between membranes, with rates that can approach those mediated by NPC2. The results suggest that sterol transfer from CD to membranes occurs by a collisional transfer mechanism involving direct interaction of CD with membranes, similar to that shown previously for NPC2. For CD, however, absolute rates are slower compared to NPC2 for a given concentration, and the lysosomal phospholipid lysobisphosphatidic acid (LBPA) does not stimulate rates of sterol transfer between membranes and CD. As expected from the apparent absence of interaction between CD and NPC2, the addition of CD to NPC2-deficient fibroblasts rapidly rescued the cholesterol accumulation phenotype. Thus, the recent observations of CD efficacy in mouse models of NPC disease are likely the result of CD enhancement of cholesterol transport between membranes, with rapid sterol transfer occurring during CD--membrane interactions.

Biophysical study of resin acid effects on phospholipid membrane structure and properties

DEFF Research Database (Denmark)

Jagalski, Vivien; Barker, Robert; Topgaard, Daniel

2016-01-01

Hydrophobic resin acids (RAs) are synthesized by conifer trees as part of their defense mechanisms. One of the functions of RAs in plant defense is suggested to be the perturbation of the cellular membrane. However, there is a vast diversity of chemical structures within this class of molecules, ...

Stability of sonicated aqueous suspensions of phospholipids under air.

Science.gov (United States)

Almog, R; Forward, R; Samsonoff, C

1991-12-01

The stability of phospholipids in liposomal aqueous suspension against oxidative degradation in air was investigated using spectrophotometric indices, glutathione peroxidase reactivity and thin layer chromatography. Zwitterionic phospholipid was found to be susceptible to degradation via oxidation of polyunsaturated hydrocarbon chains and ester hydrolysis, producing oxidized lysophosphatide and free fatty acid derivatives. These products were characterized as hydroperoxides based on their reactivity with the selenium-dependent glutathione peroxidase isolated from human erythrocytes. Lecithin in Tris buffer was more resistant to hydrolysis than in water. The sonication of 8.0 mM of soybean phosphatidylcholine (SB-PC) suspension in 0.1 M Tris (pH 7.5) in the presence of air produced relatively high concentration of conjugated diene hydroperoxide, but a small amount of hydrolyzed products. Anionic phospholipids, such as egg-phosphatidylglycerol (egg-PG), demonstrated higher resistance to air oxidation than the zwitterionic lecithin, but its oxidation was promoted by sonication.

Phosphorus starvation induces membrane remodeling and recycling in Emiliania huxleyi.

Science.gov (United States)

Shemi, Adva; Schatz, Daniella; Fredricks, Helen F; Van Mooy, Benjamin A S; Porat, Ziv; Vardi, Assaf

2016-08-01

Nutrient availability is an important factor controlling phytoplankton productivity. Phytoplankton contribute c. 50% of the global photosynthesis and possess efficient acclimation mechanisms to cope with nutrient stress. We investigate the cellular response of the bloom-forming coccolithophore Emiliania huxleyi to phosphorus (P) scarcity, which is often a limiting factor in marine ecosystems. We combined mass spectrometry, fluorescence microscopy, transmission electron microscopy (TEM) and gene expression analyses in order to assess diverse cellular features in cells exposed to P limitation and recovery. Early starvation-induced substitution of phospholipids in the cells' membranes with galacto- and betaine lipids. Lipid remodeling was rapid and reversible upon P resupply. The PI3K inhibitor wortmannin reduced phospholipid substitution, suggesting a possible involvement of PI3K- signaling in this process. In addition, P limitation enhanced the formation and acidification of membrane vesicles in the cytoplasm. Intracellular vesicles may facilitate the recycling of cytoplasmic content, which is engulfed in the vesicles and delivered to the main vacuole. Long-term starvation was characterized by a profound increase in cell size and morphological alterations in cellular ultrastructure. This study provides cellular and molecular basis for future ecophysiological assessment of natural E. huxleyi populations in oligotrophic regions. © 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.

Phospholipid-Coated Mesoporous Silica Nanoparticles Acting as Lubricating Drug Nanocarriers

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Tao Sun

2018-05-01

Full Text Available Osteoarthritis (OA is a severe disease caused by wear and inflammation of joints. In this study, phospholipid-coated mesoporous silica nanoparticles (MSNs@lip were prepared in order to treat OA at an early stage. The phospholipid layer has excellent lubrication capability in aqueous media due to the hydration lubrication mechanism, while mesoporous silica nanoparticles (MSNs act as effective drug nanocarriers. The MSNs@lip were characterized by scanning electron microscope, transmission electron microscope, Fourier transform infrared spectrum, X-ray photoelectron spectrum, thermogravimetric analysis and dynamic light scattering techniques to confirm that the phospholipid layer was coated onto the surface of MSNs successfully. A series of tribological tests were performed under different experimental conditions, and the results showed that MSNs@lip with multi-layers of phospholipids greatly reduced the friction coefficient in comparison with MSNs. Additionally, MSNs@lip demonstrated sustained drug release behavior and were biocompatible based on CCK-8 assay using MC3T3-E1 cells. The MSNs@lip developed in the present study, acting as effective lubricating drug nanocarriers, may represent a promising strategy to treat early stage OA by lubrication enhancement and drug delivery therapy.

Dynamics and energetics of the mammalian phosphatidylinositol transfer protein phospholipid exchange cycle.

Science.gov (United States)

Grabon, Aby; Orłowski, Adam; Tripathi, Ashutosh; Vuorio, Joni; Javanainen, Matti; Róg, Tomasz; Lönnfors, Max; McDermott, Mark I; Siebert, Garland; Somerharju, Pentti; Vattulainen, Ilpo; Bankaitis, Vytas A

2017-09-01

Phosphatidylinositol-transfer proteins (PITPs) regulate phosphoinositide signaling in eukaryotic cells. The defining feature of PITPs is their ability to exchange phosphatidylinositol (PtdIns) molecules between membranes, and this property is central to PITP-mediated regulation of lipid signaling. However, the details of the PITP-mediated lipid exchange cycle remain entirely obscure. Here, all-atom molecular dynamics simulations of the mammalian StART-like PtdIns/phosphatidylcholine (PtdCho) transfer protein PITPα, both on membrane bilayers and in solvated systems, informed downstream biochemical analyses that tested key aspects of the hypotheses generated by the molecular dynamics simulations. These studies provided five key insights into the PITPα lipid exchange cycle: (i) interaction of PITPα with the membrane is spontaneous and mediated by four specific protein substructures; (ii) the ability of PITPα to initiate closure around the PtdCho ligand is accompanied by loss of flexibility of two helix/loop regions, as well as of the C-terminal helix; (iii) the energy barrier of phospholipid extraction from the membrane is lowered by a network of hydrogen bonds between the lipid molecule and PITPα; (iv) the trajectory of PtdIns or PtdCho into and through the lipid-binding pocket is chaperoned by sets of PITPα residues conserved throughout the StART-like PITP family; and (v) conformational transitions in the C-terminal helix have specific functional involvements in PtdIns transfer activity. Taken together, these findings provide the first mechanistic description of key aspects of the PITPα PtdIns/PtdCho exchange cycle and offer a rationale for the high conservation of particular sets of residues across evolutionarily distant members of the metazoan StART-like PITP family. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Membrane Affinity of Platensimycin and Its Dialkylamine Analogs

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Ian Rowe

2015-08-01

Full Text Available Membrane permeability is a desired property in drug design, but there have been difficulties in quantifying the direct drug partitioning into native membranes. Platensimycin (PL is a new promising antibiotic whose biosynthetic production is costly. Six dialkylamine analogs of PL were synthesized with identical pharmacophores but different side chains; five of them were found inactive. To address the possibility that their activity is limited by the permeation step, we calculated polarity, measured surface activity and the ability to insert into the phospholipid monolayers. The partitioning of PL and the analogs into the cytoplasmic membrane of E. coli was assessed by activation curve shifts of a re-engineered mechanosensitive channel, MscS, in patch-clamp experiments. Despite predicted differences in polarity, the affinities to lipid monolayers and native membranes were comparable for most of the analogs. For PL and the di-myrtenyl analog QD-11, both carrying bulky sidechains, the affinity for the native membrane was lower than for monolayers (half-membranes, signifying that intercalation must overcome the lateral pressure of the bilayer. We conclude that the biological activity among the studied PL analogs is unlikely to be limited by their membrane permeability. We also discuss the capacity of endogenous tension-activated channels to detect asymmetric partitioning of exogenous substances into the native bacterial membrane and the different contributions to the thermodynamic force which drives permeation.

An extended study of the phase separation between phospholipids and grafted polymers on a bilayer biomembrane

Energy Technology Data Exchange (ETDEWEB)

Benhamou, M; Joudar, I; Kaidi, H; Elhasnaoui, K; Ridouane, H; Qamar, H, E-mail: benhamou.mabrouk@gmail.com [Laboratoire de Physique des Polymeres et Phenomenes Critiques, Faculte des Sciences, Ben M' sik, PO Box 7955, Casablanca (Morocco)

2011-06-01

We re-examine here the phase separation between phospholipids and adsorbed polymer chains on a fluid membrane with a change in some suitable parameter (temperature). Our purpose is to quantify the significant effects of the solvent quality and of the polydispersity of adsorbed loops formed by grafted polymer chains on the segregation phenomenon. To this end, we elaborate on a theoretical model that allows us to derive the expression for the mixing free energy. From this, we extract the phase diagram shape in the composition-temperature plane. Our main conclusion is that the polymer chain condensation is very sensitive to the solvent quality and to the polydispersity of loops of adsorbed chains.

Impact of oxidized phospholipids on the structural and dynamic organization of phospholipid membranes: a combined DSC and solid state NMR study

Czech Academy of Sciences Publication Activity Database

Wallgren, M.; Beranová, Lenka; Pham, Q. D.; Linh, K.; Lidman, M.; Procek, J.; Cyprych, K.; Kinnunen, P. K. J.; Hof, Martin; Gröbner, G.

2013-01-01

Roč. 161, DEC 2013 (2013), s. 499-513 ISSN 1359-6640 R&D Projects: GA AV ČR GEMEM/09/E006 Institutional support: RVO:61388955 Keywords : ACYL-CHAIN REVERSAL * MOLECULAR SIMULATIONS * LIPID-MEMBRANES Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 4.194, year: 2013

Simulation of water transport through a lipid membrane