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1.
Jeffrey A. Veiro Parthasarathy Nambi Lourdes L. Herold Elizabeth S. Rowe 《生物化学与生物物理学报:生物膜》1987,900(2)
We have systematically investigated the effect of short-chain n-alcohols and glycerol on the pretransition of 1,2-dipalmitoylphosphatidylcholine (DPPC) by spectrophotometry. It is found that the n-alcohols and glycerol remove the pretransition above a critical concentration for each ligand. In addition, the short-chain n-alcohols below the critical concentration decrease the pretransition temperature. The longer the aliphatic chain length of the n-alcohol (up to butanol) (a) the greater the decrease in the pretransition temperature, and (b) the lower the concentration necessary to remove the pretransition. However, glycerol differs from the short-chain n-alcohols in that it has no significant effect on either the pretransition or the main transition, but it is also capable of removing the pretransition above a critical concentration. It has previously been shown that alcohols have a biphasic effect on the main transition temperature of phosphatidylcholines (Rowe, E.S. (1983) Biochemistry 22, 3299–3305). At high alcohol concentrations, the main transition is not thermodynamically reversible (Rowe, E.S. (1985) Biochim. Biophys. Acta 813, 321–330). Recently, Simon and McIntosh (Biochim. Biophys. Acta (1984) 773, 169–172) have identified that at high ethanol concentration DPPC exists in the interdigitated phase. The critical ligand concentration at which the pretransition disappears coincides with the induction of main transition hysteresis and the biphasic alcohol effect in the main transition. These three effects appear to correlate with the induction of the interdigitated gel state by alcohols and glycerol. 相似文献
2.
The relaxation processes involved in the pretransition of dipalmitoylphosphatidylcholine (DPPC) multilamellar vesicles were investigated by the measurements of transmitted light intensity under crossed polars and freeze fracture electron microscopy. Temperature jump experiments from the ripple (Pβ′) phase into the gel (Lβ′) phase suggest that there must be at least three relaxation stages, one of which has a relaxation time much shorter than those described previously. Freeze fracture electron microscopy observations of the surface structure during the relaxation process reveals that the defects in the ripple pattern, i.e. the open terminal ends of each ripple ridge line, may be the primary origin of the growth of the Lβ′ phase, and that the phase conversion may take place exclusively at the pointed end of a ripple ridge line. Thus the Lβ′ phase grows almost one-dimensionally between ripple structures. 相似文献
3.
Liposomes of dipalmitoylphosphatidylcholine (DPPC) prepared in increasing glycerol/glucose ratios show an increase in the absorbance at 570 nm of merocyanine spectra at temperatures below the phase transition. Since this effect is not observed when liposomes are prepared in solutions containing solely glucose, it is attributed to specific interactions of glycerol with the membrane phase. The increase in the 570 nm absorbance is ascribed to a partial fluidification of the membrane interface and is dependent on the distribution of the dye between the inner and the outer compartments of the liposomes and on their osmotic state. The greatest differences in the absorbance ratio are obtained when merocyanine is added to the external media. In consequence, the changes in the spectra of MC are dependent on the surface state of the liposomes which can be modified by an increase of glycerol or glucose in the external media. The present results are examined in the light of the perturbations that glycerol can induce on the barrier properties of the bilayer. 相似文献
4.
The correlation between electrotransformation and electropermeability was studied in yeast cells following the modification of their membranes by treatment with n-alcohols. It was found that the number of transformed cells decreased with increase of chain length of the alcohols used as follows: methanoln-alcohol treatment. The lack of a unidirectional link between permeability and transformation leads to the assumption that the mechanism of DNA introduction into the cell could not be interpreted solely as a result of the existence of pores.
Correspondence to: I. Tsoneva 相似文献
5.
Elizabeth S. Rowe 《生物化学与生物物理学报:生物膜》1982,685(2):105-108
The gel to liquid crystal phase transition of dipalmitoylphosphatidylcholine (DPPC) has been followed by the change in absorbance at 400 nm; this change is due to the change in lipid light scattering properties during the transition. The effect of sucrose on the change in absorbance during the transition of DPPC has been investigated. It has been shown that the presence of sucrose or glycerol in the multilamellar liposome suspension increases the change in absorbance due to the main transition, decreases the total absorbance, and decreases the change in absorbance due to the pretransition. This effect of sucrose and glycerol is shown to be an optical effect which is correlated with solvent index of refraction. 相似文献
6.
LUSH is an odorant binding protein expressed in the olfactory organs of Drosophila melanogaster that is required for the detection of alcohol in adult flies. Here we demonstrate that, in the absence of ligand, in vitro LUSH exists in a partial molten globule state. The presence of short-chain n-alcohols at pharmacologically relevant concentrations less than 50 mM shifts the conformational equilibrium to a more compact state that exhibits reduced binding of the fluorescent dye 1-anilino-8-naphthalenesulfonic acid. Equilibrium unfolding studies of LUSH-alcohol complexes reveal that, for a series of short-chain n-alcohols, each methylene group can contribute approximately 1 K cal mol(-1) to the overall stability of the protein-alcohol complex. Using NMR spectroscopy, we have identified the regions of LUSH that show increased conformational stability on binding alcohols. These residues primarily line the alcohol-binding pocket. The results presented here provide a direct measure of the degree of stability that alcohol imparts on LUSH. These observations may represent a model for how ethanol can stabilize alternative protein conformations in alcohol-sensitive human proteins and ultimately lead to the observed changes in higher order function throughout the central nervous system. 相似文献
7.
Effect of glycosaminoglycans and divalent cations on the thermal properties of dipalmitoylphosphatidylcholine 总被引:1,自引:1,他引:0
The effect of glycosaminoglycans (GAG) and divalent cations on the thermal properties of dipalmitoyl-phosphatidylcholine (DPPC)-water systems was examined in order to model some interactions taking place on low density lipoprotein (LDL) surfaces. The thermal properties of these systems were measured by differential scanning calorimetry (DSC). According to the results, all three glycosaminoglycans used (chondroitin-4-sulfate, chondroitin-6-sulfate and heparin) were effective but to a different extent. Calcium ions enhance the interaction more than magnesium ions, probably because divalent cations form bridges between the negatively charged groups of GAGs and the headgroups of lipids. It is conceivable that similar processes might occur in the case of LDL. 相似文献
8.
Phospholipids with sites of unsaturation are targets of peroxidation. We investigated the effect of two types of lipids with identical headgroups, sphingomyelins (SMs) and dipalmitoylphosphatidylcholine (DPPC), on the extent of oxidation of stearoyl-arachidonoyl phosphatidylcholine (SAPC) with four double bonds. Peroxidation was induced with tert-butylhydroperoxide and FeCl(2) at 35 degrees C. The decrease of SAPC versus DPPC, or N-palmitoyl SM, or N-stearoyl SM, was monitored by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) at various reaction times. For corresponding molar ratios of DPPC:SAPC and SM:SAPC, SAPC oxidized faster and to a greater extent when DPPC, rather than N-palmitoyl SM or bovine brain SMs, was present. However, at 35 degrees C the hydrophobic tails in SM mixtures were more disordered than in those of DPPC. The slower oxidation of SAPC in SM-rich vesicles may result from the presence of a tight network of H-bonds that bridge neighboring SM molecules and poses a stronger interfacial barrier to the passage of oxidants. 相似文献
9.
Oxysterols are oxygenated derivatives of cholesterol that form a class of potent regulatory molecules with diverse biological activity. Given the implications of oxysterols in several physiological/pathophysiological pathways of human diseases, it is important to identify how their presence affects the biophysical properties of cell membranes. In this article we first describe the structure, formation, and biological functions of oxysterols, and previous work on the effect of these molecules on the structure and phase behavior of lipid membranes. We then present results of our X-ray diffraction experiments on aligned multilayers of dipalmitoylphosphatidylcholine (DPPC) membranes containing ring-substituted oxysterols. The effect of these molecules on the phase behavior of DPPC membranes is found to be very similar to that of cholesterol. All the oxysterols studied induce a modulated phase in DPPC membranes, similar to that reported in DPPC–cholesterol membranes. However, some differences are observed in the ability of these molecules to suppress the main transition of the lipid and to induce chain ordering, which might be related to differences in their orientation in the bilayer. 相似文献
10.
Effect of trehalose on the phase properties of hydrated and lyophilized dipalmitoylphosphatidylcholine multilayers 总被引:3,自引:0,他引:3
The structure and thermal behavior of hydrated and lyophilized dipalmitoylphosphatidylcholine (DPPC) multilayers in the presence of trehalose were investigated by differential scanning calorimetry and X-ray diffraction methods. Trehalose enters the aqueous space between hydrated bilayers and increases the interbilayer separation (from 0.36 to 1.37 nm in the different DPPC phases at 1 M trehalose). It does not affect the lipid chain packing and also the slow isothermal conversion at 4 degrees C of the metastable L beta' phase into the equilibrium crystalline Lc phase. Addition of trehalose leads to a slight upward shift (about 1 degrees C at 1 M trehalose) of the three phase transitions (sub-, pre-, and main transition) in fully hydrated DPPC while their other properties (enthalpy, excess specific heat, and transition width) remain unchanged. The effect of trehalose on the thermal behavior of DPPC multilayers freeze-dried from an initially completely hydrated state is qualitatively similar to that of water. These data support the "water replacement" hypothesis about trehalose action. It is suggested that trehalose prevents the formation of direct interbilayer hydrogen bonds in states of low hydration. 相似文献
11.
Effect of dipalmitoylphosphatidylcholine vesicle curvature on the reaction with human apolipoprotein A-I 总被引:1,自引:0,他引:1
Large unilamellar vesicles of dipalmitoylphosphatidylcholine (DPPC) were prepared by sonication and were fractionated by gel filtration on Sepharose Cl-2B in the size range from 180- to 380-A Stokes radii. Negatively stained electron micrographs of these preparations indicated the presence of unilamellar, spheroidal structures of the expected size. Fluorescence polarization of diphenylhexatriene, dissolved in the vesicles, revealed progressively broader phase transitions, shifted to lower temperatures for vesicles of decreasing sizes. The fractionated unilamellar vesicles and multilamellar vesicles of DPPC were reacted with human apolipoprotein A-I at 41 degrees C for periods from 1 to 120 h. The reaction mixtures were then passed through a Bio-Gel A-5m column to separate unreacted lipid vesicles and protein from micellar complexes of DPPC with apolipoprotein A-I. Smaller vesicles were much more reactive than larger vesicles or multilamellar vesicles with the apolipoprotein. This difference in reactivity was explained by the increasing bilayer curvature of smaller vesicles which changes the packing of DPPC molecules in the bilayer and facilitates its penetration by the apolipoprotein. 相似文献
12.
P. Maček Marialucia Zecchini Katja Stanek Gianfranco Menestrina 《European biophysics journal : EBJ》1997,25(3):155-162
Equinatoxin II, a 19.8 kDa pore-forming toxin from the sea anemone Actinia equina, was examined for hemolytic activity and permeabilization of small unilamellar lipid vesicles (SUV) in the presence of increasing
amounts of n-alcohols (methanol to n-octanol) and fatty acids (palmitic and palmitoleic acid). We observed an enhancement
of toxin activity which was dependent on the concentration of the membrane partitioning additive. An exception was palmitic
acid which exerted a bimodal role. While at low bulk concentrations it increased toxin-induced hemolysis, above 3 μM bulk
concentration it was inhibitory; in neither case was it efficient in promoting release of the fluorescent marker calcein from
SUV. The increased permeabilization activity was correlated with an increase in the amount of toxin bound as indicated by
changes in the intrinsic toxin fluorescence. In the case of n-alcohols, at least, these effects appeared to depend on the
actual amount of alcohol present inside the membrane rather than on its specific chemical nature. This suggests that the observed
effects could be due to changes of the biophysical properties of the lipid bilayer, such as thickness, lipid acyl-chain ordering,
and dielectric constant induced by the partitioned additives.
Received: 27 March 1996 / Accepted: 10 October 1996 相似文献
13.
Effect of glycerol and dihydroxyacetone on hepatic lipogenesis 总被引:2,自引:0,他引:2
Glycerol is a dietary component which is metabolized primarily by the liver and kidney where it is used mainly for glucose synthesis. The metabolism of glycerol is very similar to that of dihydroxyacetone which can be considered its more oxidized counterpart. The effects of these substrates on hepatic lipogenesis and gluconeogenesis were examined. In isolated hepatocytes, 10 mM dihydroxyacetone caused a large increase in glucose output and stimulated lipogenesis without affecting the lactate/pyruvate ratio or the total ATP content of the cells. (As compared to dihydroxyacetone, 10 mM glycerol was less effective as a gluconeogenic substrate, increased the lactate/pyruvate ratio, caused a slight decrease in the total ATP content, and inhibited lipogenesis by at least 40% depending on the type of diet fed to the rats.) The fall in ATP levels was very small and did not correlate with the changes in fatty acid synthesis. The immediate cause of the inhibition of lipogenesis, brought about by glycerol in hepatocytes from sucrose fed rats, seemed to be a large decrease in pyruvate levels. This did not result from impairment of glycolysis but from a rise in the cytosolic NADH/NAD ratio. 相似文献
14.
Norman B. Smith 《Chemistry and physics of lipids》1981,29(3):277-282
Sonicated dispersions of 1,2-dipalmitoylsn-glycero-3-phosphorylcholine and of 1,3-dipalmitoylglycero-2-phosphorylcholine were examined by proton nuclear magnetic resonance (NMR) as a function of temperature. The —(CH2)n)— peak in the spectrum of the sn-3-isomer of dipalmitoylphosphatidylcholine showed the characteristic dramatic changes in the peak intensity and width associated with the phase transition between the liquid crystalline and gel states of the phospholipid. This occurred over a 2–3°C temperature range with the midpoint of the transition at 38.5°C. With the 2-isomer the change in phase took place over a similar temperature range but the midpoint was at 33.8°C. This lower phase transition temperature is presumably the result of increased acyl chain mobility caused by the increased separation of the two acyl chains by the centre carbon of the glycerol backbone. The effect of sonication of the broadening of the range and lowering of the midpoint temperature of the phase transition from that of the corresponding unsonicated dispersions was similar with each isomer. This suggests that the overall geometry of the sonicated vesicles of the isomers is similar. 相似文献
15.
Effect of stereoconfiguration on ripple phases (P beta') of dipalmitoylphosphatidylcholine 总被引:1,自引:0,他引:1
J A Zasadzinski 《Biochimica et biophysica acta》1988,946(2):235-243
Mixtures of sn-1 (D) and sn-3 (L) enantiomers of fully hydrated dipalmitoylphosphatidylcholine (DPPC) were studied with differential scanning calorimetry and freeze-fracture microscopy. The pretransition temperature of racemic mixtures of DPPC was 1.8 C degrees below that of either pure sn-1 or sn-3 enantiomers, which had similar pretransition temperatures. The main transition temperature of racemic mixtures was also depressed, but to a lesser extent, 0.8 C degrees. Freeze-fracture images of liposomes of sn-1, sn-3, and racemic mixtures of DPPC frozen from the P beta' phase showed well-defined ripples of wavelength 13 nm. Lipid stereoconfiguration had no effect on ripple wavelength, configuration or amplitude, or on the number and nature of surface defects. 相似文献
16.
17.
Single-molecule spectroscopy at cryogenic temperatures was used to examine the impact of buffer solution, glycerol/buffer mixtures (25% and 66%), and poly(vinyl alcohol) (PVA) films on the conformation of photosystem I (PSI) from Thermosynechoccocus elongatus. PSI holds a number of chromophores embedded at different places within the protein complex that show distinguishable fluorescence at low temperatures. The fluorescence emission from individual complexes shows inter- and intracomplex heterogeneity depending on the solution wherein PSI was dissolved. Statistical evaluation of spectra of a large number of complexes shows that the fluorescence emission of some of these chromophores can be used as sensors for their local nanoenvironment and some as probe for the conformation of the whole protein complex. Preparation in glycerol/buffer mixtures yields a high homogeneity for all chromophores, indicating a more compact protein conformation with less structural variability. In buffer solution a distinct heterogeneity of the chromophores is observed. PSI complexes in PVA show highly heterogeneous spectra as well as a remarkable blue shift of the fluorescence emission, indicating a destabilization of the protein complex. Photosystem I prepared in PVA cannot be considered fully functional, and conclusions drawn from experiments with PSI in PVA films are of questionable value. 相似文献
18.
19.
The effects of n-alcohols on sarcoplasmic reticulum vesicles 总被引:1,自引:0,他引:1
20.
The effect of 1-anilinonaphthalene-8-sulfonate (ANS) on the thermotropic phase transition of dipalmitoylphosphatidylcholine (DPPC) bilayers was examined by differential scanning calorimetry. The main phase transition temperature was found to be shifted to lower values in the presence of the probe. The shift strongly depends on pH and the presence of salts. This indicates that the penetration of the probe of the hydrocarbon moiety of the bilayer is influenced by coulombic interactions. Pretransition phenomena are also affected. The implications for the interpretation of experimental data of biomembrane studies are discussed. 相似文献