| For medical practitioners and the general public - Cholesterol Journal Article Catalog. |
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| Interaction of cholesterol with sphingosine: physicochemical characterization and impact on intestinal absorption Garmy, N., N. Taieb, et al. (2005), J Lipid Res 46(1): 36-45. Abstract: Molecular associations between sphingomyelin and cholesterol provide a molecular basis for the colocalization of these lipids in plasma membrane microdomains (lipid rafts) and for the inhibitory effect of sphingomyelin on the intestinal absorption of cholesterol. Using surface pressure measurements at the air-water interface, we showed that sphingosine, the common sphingoid backbone of most sphingolipids, formed condensed lipid complexes with cholesterol. Structure-activity relationship studies with long-chain analogs of sphingosine, together with molecular mechanics simulations, were consistent with a specific interaction between sphingosine and the alpha face of cholesterol. The uptake of micellar cholesterol and the effect of sphingosine on cholesterol absorption were studied with two human model intestinal epithelial cell lines, Caco-2 and HT-29-D4. Real-time PCR quantifications of the putative cholesterol transporter Niemann-Pick C1 like 1 (NPC1L1) mRNA revealed that, in these cell lines, the activity of cholesterol transport correlated with the level of NPC1L1 expression. In both cell lines, sphingosine induced a dose-dependent decrease of cholesterol absorption. Yet the effect of sphingosine was more dramatic in Caco-2 cells, which also displayed the highest expression of NPC1L1 mRNA. Altogether, these data suggested that sphingosine interacts specifically with cholesterol and inhibits the intestinal NPC1L1-dependent transport of micellar cholesterol. |
| Interaction of cholesterol with synthetic sphingomyelin derivatives in mixed monolayers Gronberg, L., Z. S. Ruan, et al. (1991), Biochemistry 30(44): 10746-54. Abstract: To study the structural requirements of the molecular interactions between cholesterol and sphingomyelins in model membranes, sphingomyelin derivatives were synthesized in which (a) the 3-hydroxy group was replaced with a hydrogen atom or with a methoxy, ethoxy, or tetrahydropyranyloxy group, (b) the N-acyl chain length was varied, and (c) the N-acyl chain length contained an alpha-hydroxy group. The chemical syntheses of these derivatives from DL-erythro-sphingosine are reported. The properties of these sphingomyelin derivatives were examined in monolayer membranes at the air/water interface. The mean molecular area of the pure N-stearoylsphingomyelin derivatives was determined, and the effects of cholesterol on the condensation of sphingomyelin packing in the monolayer were recorded. It was observed that replacement of the 3-hydroxy group of sphingomyelin with a hydrogen atom or its substitution with a methoxy or ethoxy group did not affect the ability of cholesterol to condense the molecular packing in monolayers. Even when a bulky tetrahydropyranyloxy group was introduced at the 3-hydroxy position of egg sphingomyelin, cholesterol was still able to condense the molecular packing of this derivative. The condensing effect of cholesterol on derivatives of N-stearoyl-SPMs was significantly larger than the comparable effect observed with 1,2-distearoyl-sn-glycero-3-phosphocholine or 1,2-dipalmitoyl-sn-glycero-3-phosphocholine. Our results with 3-hydroxysphingomyelins having differing N-acyl chain lengths (i.e., N-stearoyl, N-myristoyl, and N-lauroyl), and with 3-hydroxy-N-(alpha-hydroxypalmitoyl)sphingomyelin also indicated that cholesterol was able to induce condensation of the molecular packing.(ABSTRACT TRUNCATED AT 250 WORDS) |
| Interaction of cholesterol with various glycerophospholipids and sphingomyelin Sankaram, M. B. and T. E. Thompson (1990), Biochemistry 29(47): 10670-5. Abstract: The influence of cholesterol on the phase behavior of glycerophospholipids and sphingomyelins was investigated by spin-label electron spin resonance (ESR) spectroscopy. 4-(4,4-Dimethyl-3-oxy-2-tridecyl-2-oxazolidinyl)butanoic acid (5-SASL) and 1-stearoyl-2-4-(4,4-dimethyl-3-oxy-2-tridecyl-2-oxazolidinyl)butanoy l-sn- glycero-3-phosphocholine (5-PCSL) spin-labels were employed for this purpose. The outer hyperfine splitting constants, Amax, measured from the spin-label ESR spectra as a function of temperature were taken as empirical indicators of cholesterol-induced changes in the acyl chain motions in the fluid state. The Amax values of 5-PCSL exhibit a triphasic dependence on the concentration of cholesterol for phosphatidylcholines and bovine brain sphingomyelin. We interpret this dependence as reflecting the existence of liquid-disordered, ld, liquid-ordered, lo, and coexistence regions, ld + lo. The phase boundary between the ld and the two-phase region and the boundary between the lo and the two-phase region in the phosphatidylcholine-cholesterol systems coalesce at temperatures 25-33 degrees C above the main-chain melting transition temperature of the cholesterol-free phosphatidylcholine bilayers. In the case of bovine brain sphingomyelin, the ld-lo phase coalescence occurs about 47 degrees C above the melting temperature of the pure sphingomyelin. The selectivity of interaction of cholesterol with glycerophospholipids of varying headgroup charge was studied by comparing the cholesterol-induced changes in the Amax values of derivatives of phosphatidylcholine, phosphatidic acid, phosphatidylethanolamine, phosphatidylglycerol, and phosphatidylserine spin-labeled at the fifth position of the sn-2 chain.(ABSTRACT TRUNCATED AT 250 WORDS) |
| Interaction of cholesterol-conjugated alkylating oligonucleotide derivatives with cellular biopolymers Ryte, A. S., V. N. Karamyshev, et al. (1992), FEBS Lett 299(2): 124-6. Abstract: Interactions of oligonucleotide derivatives with mammalian cells and cellular biopolymers have been investigated. The derivatives were oligonucleotides bearing an alkylating 2-chloroethylamino group at the 3'-end and a cholesterol residue at the 5'-terminal phosphate. These compounds are readily taken up by cells and react with cellular DNA, RNA and some proteins which may play a role in delivery of the compounds into cells. |
| Interaction of cholesterol-crystallization-promoting proteins with vesicles de Bruijn, M. A., B. G. Goldhoorn, et al. (1995), Biochem J 305 (Pt 1): 93-6. Abstract: In this study, the interaction of mucin and concanavalin A-binding proteins isolated from human bile with cholesterol/phospholipid vesicles was investigated. Using resonance energy transfer assays originally developed by Struck, Hoekstra and Pagano (1981) Biochemistry 20, 4093-4099, no significant protein-induced fusion or aggregation of vesicles was demonstrated. Instead of fusion, these proteins induced destabilization of cholesterol/phospholipid vesicles, as monitored by release of entrapped carboxyfluorescein. A good correlation (rho = 0.81) was obtained between the extent of leakage and the nucleation-promoting activity of the concanavalin A-binding proteins. We conclude that aggregation or fusion of cholesterol/phospholipid vesicles is not an obligatory step in cholesterol crystallization. Biliary protein-induced crystallization seems to be preceded by vesicle disruption. |
| Interaction of dietary fat saturation and cholesterol level on cholesterol synthesis measured using deuterium incorporation Jones, P. J., A. H. Lichtenstein, et al. (1994), J Lipid Res 35(6): 1093-101. Abstract: To examine interactive effects of dietary fat saturation and cholesterol level on serum lipids and de novo cholesterogenesis, moderately hypercholesterolemic subjects were fed solid-foods diets containing 30% fat (80 mg C.1000 Kcal-1) in which 2/3 fat was either corn oil or beef tallow, with and without 120 mg C.1000 Kcal-1, for 5 wk. At the end of each diet period, subjects were given deuterium (D) oxide orally and de novo cholesterol synthesis was measured over 24 h from D incorporation into cholesterol as fractional synthesis rates (FSR) and absolute synthetic rate (ASR) into the rapid exchangeable cholesterol pool. Plasma total and low density lipoprotein levels were elevated (P < 0.01) with both beef tallow and cholesterol feeding. High density lipoprotein (HDL) levels were not influenced by fat saturation or cholesterol level; however, HDL was higher with addition of cholesterol to corn oil versus beef tallow (P < 0.02). Plasma triglycerides were higher (P < 0.02) with beef tallow feeding but were not influenced by cholesterol level. FSR was increased (P < 0.02) by feeding corn oil, versus beef fat, but not by dietary cholesterol level. Calculated cholesterol pools sizes did not differ across groups; however, ASR was also elevated with corn oil versus beef tallow feeding (P < 0.02). Results indicate that corn oil feeding lowers circulating cholesterol by mechanisms other than reduced synthesis, and that cholesterol at the level of supplementation used is not associated with feedback inhibition of cholesterogenesis. However, with the exception of HDL levels, dietary fat saturation and cholesterol levels do not interactively influence circulating lipoprotein cholesterol levels and cholesterol synthesis. |
| Interaction of ethanol, cholesterol and vitamin A on phospholipid methylation in hepatic microsomes of rats Srivastava, R., N. Srivastava, et al. (1990), Int J Vitam Nutr Res 60(3): 236-9. Abstract: Effect of ethanol, cholesterol and vitamin A either alone or in combination has been studied on cyt P450 and phospholipid methylation in hepatic microsomes of rats. Ethanol increased cyt P450 activity and decreased phospholipid methylation. Cholesterol effects were opposite to ethanol. Combination of cholesterol and ethanol decreased cyt P450 activity and increased phospholipid methylation. Vitamin A, which alone did not affect cyt P450 activity and phospholipid methylation behaved like cholesterol when given along with ethanol. |
| Interaction of free apolipoproteins with macrophages. Formation of high density lipoprotein-like lipoproteins and reduction of cellular cholesterol Hara, H. and S. Yokoyama (1991), J Biol Chem 266(5): 3080-6. Abstract: Mouse peritoneal macrophages, loaded with cholesteryl ester by incubating with acetylated human low density lipoprotein containing 3Hcholesteryl oleate, were exposed to purified human apolipoproteins (apo) A-I, A-II, C-III, or E in aqueous solutions. Unesterified cholesterol was released into the medium in the presence of apoA-I, -A-II, or -E, accompanied by the decrease in intracellular cholesteryl ester. ApoC-III had no such effects. Apparent Km values of the cholesterol release were estimated as 0.11, 0.14, and 0.24 microM, and Vmax values 35, 11, and 14 micrograms of cholesterol/mg of cell protein/6 h, for apoA-I, -A-II, and -E, respectively. The products formed with apoA-I, -A-II, or -E in the media were analyzed by density gradient ultracentrifugation when the cells were preloaded with 3Hcholesteryl oleate-acetylated low density lipoproteins and 3Hcholine. Free 3Hcholesterol, 3Hphosphatidylcholine, and 3Hsphingomyelin were detected coincidentally as a symmetric peak at the density of 1.1 in each case. In the complex of lipids and apoA-I or apoA-II, the weight ratios of apolipoprotein/cholesterol/phosphatidylcholine/sphingomyelin/lysophosphati dyl- choline were estimated as 2.2:1:0.6:0.2:0.07 and 4.0:1:0.5:0.3:0.07, respectively. Both of the products formed with apoA-I and -A-II migrated slower than plasma high density lipoprotein in electrophoresis on agarose gel. Because the Km values are as low as 1:340-400, 1:140-160, and 1:6-8 of plasma concentrations of apoA-I, -A-II, and -E, respectively, the results have physiological relevance for a function of the free apolipoproteins in interstitial fluid to form high density lipoprotein and to reduce cellular cholesterol. |
| Interaction of heptakis (2,3,6-tri-O-methyl)-beta-cyclodextrin with cholesterol in aqueous solution Nishijo, J., S. Moriyama, et al. (2004), Chem Pharm Bull (Tokyo) 52(12): 1405-10. Abstract: The interaction of cholesterol with heptakis (2,3,6-tri-O-methyl)-beta-cyclodextrin (TOM-beta-CyD) was investigated in water using solubility method. It was found that TOM-beta-CyD forms two kinds of soluble complexes, with molar ratios of 1:1 and 1:2 (cholesterol:TOM-beta-CyD). The thermodynamic parameters for 1:1 and 1:2 complex formation of cholesterol with TOM-beta-CyD were: DeltaG0(1:1)=-11.0 kJ/mol at 25 degrees C (K1:1=7.70 x 10 M(-1)); DeltaH0(1:1)=-1.28 kJ/mol; TDeltaS0(1:1)=9.48 kJ/mol; DeltaG0(1:2)=-27.8 kJ/mol at 25 degrees C (K1:2)=7.55 x 10(4) M(-1)); DeltaH0(1:2)=-0.57 kJ/mol; TDeltaS0(1:1)=27.3 kJ/mol. The formation of the 1:2 complex occurred much more easily than that of the 1:1 complex. The driving force for 1:1 and 1:2 complex formation was suggested to be exclusively hydrophobic interaction. Based on the measurements of proton nuclear magnetic resonance spectra and studies with Corey-Pauling-Koltun atomic models, the probable structures of the 1:2 complex were estimated. In addition, the interaction of TOM-beta-CyD with cholesterol was compared with that of heptakis (2,6-di-O-methyl)-beta-CyD (DOM-beta-CyD). The interaction of TOM-beta-CyD is more hydrophobic than that of DOM-beta-CyD, and the life time of the complexed TOM-beta-CyD is sufficiently long to give separated signals, at the NMR time scale, which differs from that of complexed DOM-beta-CyD. |
| Interaction of hormone-sensitive lipase with steroidogenic acute regulatory protein: facilitation of cholesterol transfer in adrenal Shen, W. J., S. Patel, et al. (2003), J Biol Chem 278(44): 43870-6. Abstract: Hormone-sensitive lipase (HSL) is responsible for the neutral cholesteryl ester hydrolase activity in steroidogenic tissues. Through its action, HSL is involved in regulating intracellular cholesterol metabolism and making unesterified cholesterol available for steroid hormone production. Steroidogenic acute regulatory protein (StAR) facilitates the movement of cholesterol from the outer mitochondrial membrane to the inner mitochondrial membrane and is a critical regulatory step in steroidogenesis. In the current studies we demonstrate a direct interaction of HSL with StAR using in vitro glutathione S-transferase pull-down experiments. The 37-kDa StAR is coimmunoprecipitated with HSL from adrenals of animals treated with ACTH. Deletional mutations show that HSL interacts with the N-terminal as well as a central region of StAR. Coexpression of HSL and StAR in Chinese hamster ovary cells results in higher cholesteryl ester hydrolytic activity of HSL. Transient overexpression of HSL in Y1 adrenocortical cells increases mitochondrial cholesterol content under conditions in which StAR is induced. It is proposed that the interaction of HSL with StAR in cytosol increases the hydrolytic activity of HSL and that together HSL and StAR facilitate cholesterol movement from lipid droplets to mitochondria for steroidogenesis. |
| Interaction of influenza virus haemagglutinin with sphingolipid-cholesterol membrane domains via its transmembrane domain Scheiffele, P., M. G. Roth, et al. (1997), Embo J 16(18): 5501-8. Abstract: Sphingolipid-cholesterol rafts are microdomains in biological membranes with liquid-ordered phase properties which are implicated in membrane traffic and signalling events. We have used influenza virus haemagglutinin (HA) as a model protein to analyse the interaction of transmembrane proteins with these microdomains. Here we demonstrate that raft association is an intrinsic property encoded in the protein. Mutant HA molecules with foreign transmembrane domain (TMD) sequences lose their ability to associate with the lipid microdomains, and mutations in the HA TMD reveal a requirement for hydrophobic residues in contact with the exoplasmic leaflet of the membrane. We also provide experimental evidence that cholesterol is critically required for association of proteins with lipid rafts. Our data suggest that the binding to specific membrane domains can be encoded in transmembrane proteins and that this information will be used for polarized sorting and signal transduction processes. |
| Interaction of lipoproteins with type II pneumocytes in vitro: morphological studies, uptake kinetics and secretion rate of cholesterol Guthmann, F., B. Harrach-Ruprecht, et al. (1997), Eur J Cell Biol 74(2): 197-207. Abstract: Apart from dipalmitoyl phosphatidylcholine, cholesterol is the most abundant surfactant lipid. About 90 to 99% of cholesterol of the alveolar surfactant is derived from serum lipoproteins. The aim of this study was to identify the lipoprotein which preferentially supplements type II pneumocytes with cholesterol destined for surfactant production. Ultrastructural investigations revealed that type II pneumocytes bind and take up HDL, LDL and VLDL. Binding and uptake of VLDL occurred even in the presence of excess LDL indicating that, besides LDL receptors, type II pneumocytes express additional binding sites for VLDL. Type II pneumocytes in primary culture are able to take up cholesterol added in the form of HDL, LDL and VLDL. Cholesterol uptake was lowest from HDL and highest from VLDL. The maximal velocity of cholesterol uptake from VLDL was more than three times that of cholesterol uptake from LDL. The half-maximal saturation of cholesterol uptake from VLDL was nearly half that of LDL. From these kinetic data and the distribution of free cholesterol among the serum lipoproteins, we calculated that the cholesterol uptake from VLDL is more than three times that of cholesterol uptake from LDL. In double-labeling experiments type II pneumocytes secreted palmitic acid-labeled phospholipids together with labeled free cholesterol taken up from lipoproteins. The secretion rates of both phospholipids and free cholesterol were stimulated to nearly the same extent by isoproterenol. From our results we conclude that type II pneumocytes interact specifically with HDL, LDL and VLDL. Cholesterol taken up in the form of the individual lipoproteins shows no difference in its availability for the formation of cholesterol ester and surfactant by type II pneumocytes in vitro. Based on the kinetic studies, it appears that VLDL is the major gateway through which cholesterol is provided to satisfy the cholesterol requirements of type II pneumocytes for the synthesis of surfactant. |
| Interaction of liposomes composed of phospholipids, GM1 ganglioside and cholesterol with human keratinocytes in culture Pitto, M., P. Palestini, et al. (1999), Arch Dermatol Res 291(4): 232-7. Abstract: We studied the possibility of supplementing human keratinocytes with exogenous lipids (phospholipids, sphingolipids and cholesterol) and evaluated their influence on cell proliferation, using cells cultured in vitro. Experiments carried out with liposomes composed of cholesterol/GM1 ganglioside and different phospholipids (5:1.5:10, M/M/M), showed that liposomes associated with cells more efficiently when they contained soya lecithin. The treatment with liposomes made of the ternary mixture did not modify the rate of cell proliferation, as assessed by the incorporation of 3H-thymidine. In contrast, the proliferation rate strongly decreased (65% with respect to the control) using the same liposomes without GM1. Experiments carried out with GM1 alone showed a strong stimulation of the proliferation rate (144% with respect to the control). Fluorescence dequenching experiments, carried out with the probe octadecyl rhodamine B chloride, showed that fusion was the main mechanism of liposome-cell interaction. Metabolic studies established that exogenously administered GM1--either embedded in liposomes or as a pure glycolipid dispersion--led to the production of several products, including ceramide. Altogether, these results show that different, opposing effects can be exerted on cell proliferation by the administration of lipids, separately or in mixtures, to human keratinocytes, and indicate the importance of a correct formulation for supplementing human keratinocytes with exogenous lipids. |
| Interaction of melittin with membrane cholesterol: a fluorescence approach Raghuraman, H. and A. Chattopadhyay (2004), Biophys J 87(4): 2419-32. Abstract: We have monitored the organization and dynamics of the hemolytic peptide melittin in membranes containing cholesterol by utilizing the intrinsic fluorescence properties of its functionally important sole tryptophan residue and circular dichroism spectroscopy. The significance of this study is based on the fact that the natural target for melittin is the erythrocyte membrane, which contains high amounts of cholesterol. Our results show that the presence of cholesterol inhibits melittin-induced leakage of lipid vesicles and the extent of inhibition appears to be dependent on the concentration of membrane cholesterol. The presence of cholesterol is also shown to reduce binding of melittin to membranes. Our results show that fluorescence parameters such as intensity, emission maximum, and lifetime of membrane-bound melittin indicate a change in polarity in the immediate vicinity of the tryptophan residue probably due to increased water penetration in presence of cholesterol. This is supported by results from fluorescence quenching experiments using acrylamide as the quencher. Membrane penetration depth analysis by the parallax method shows that the melittin tryptophan is localized at a relatively shallow depth in membranes containing cholesterol. Analysis of energy transfer results using melittin tryptophan (donor) and dehydroergosterol (acceptor) indicates that dehydroergosterol is not randomly distributed and is preferentially localized around the tryptophan residue of membrane-bound melittin, even at the low concentrations used. Taken together, our results are relevant in understanding the interaction of melittin with membranes in general, and with cholesterol-containing membranes in particular, with possible relevance to its interaction with the erythrocyte membrane. |
| Interaction of mucin with cholesterol enriched vesicles: role of mucin structural domains Afdhal, N. H., X. Cao, et al. (2004), Biomacromolecules 5(2): 269-75. Abstract: We utilized fluorescence recovery after photobleaching (FRAP) and fluorescence correlation spectroscopy (FCS) to examine the role of gallbladder mucin (GBM) in promoting the aggregation and/or fusion of cholesterol enriched vesicles. By fluorescent labeling either the vesicle or the mucin, we could examine the change in vesicle size as well as changes in mucin's diffusion constant. Both FRAP and FCS show that GBM has a profound effect in inducing vesicles to aggregate/fuse, particularly after overnight incubation. GBM mucin domains (either protease digested or reduced GBM) are not as effective as native GBM. Intact GBM alone was able to shorten crystal appearance time and increase the number of crystals nucleated by polarized optical microscopy. In summary, our findings would suggest that both glycosylated and nonglycosylated domains of GBM are involved in early aggregation of cholesterol enriched vesicles but that this effect is reversible in the absence of nonglycosylated domains. |
| Interaction of oxidized HDLs with J774-A1 macrophages causes intracellular accumulation of unesterified cholesterol Musanti, R. and G. Ghiselli (1993), Arterioscler Thromb 13(9): 1334-45. Abstract: Uptake of modified lipoproteins by resident arterial monocytes/macrophages is believed to be a key event in the formation of foam cells and thus in the early phases of atherosclerosis. Low-density lipoproteins (LDLs) that undergo oxidative changes become suitable for uptake by macrophages through a specific scavenger receptor that leads to cholesteryl ester accumulation. Because the interaction of other oxidized lipoproteins with macrophages has been poorly investigated, we studied the effect of oxidatively modified high-density lipoproteins (HDLs) on the sterol metabolism of J774-A1 macrophages. Unlike native HDLs, oxidized HDLs caused a concentration-dependent accumulation of unesterified cholesterol and decreased 14Coleate incorporation into steryl esters. Oxidized HDLs also decreased 14Cacetate incorporation into newly synthesized sterols. Cell surface binding of 125I-oxidized HDLs to the macrophages was saturable, with an apparent dissociation constant (Kd) of 0.96 nmol/mL. Both oxidized and acetylated LDLs but not native lipoproteins could compete for binding of 125I-oxidized HDL. The data support the conclusion that the effects elicited by oxidized HDLs on the sterol metabolism of macrophages are significantly different from those of native HDLs. The binding of oxidized HDLs to macrophages occurs at sites that are likely the same as those for modified LDLs. We speculate that, if occurring in vivo, HDL oxidation would generate modified lipoproteins capable of modulating the cholesterol homeostasis of macrophages. |
| Interaction of polyethyleneglycol-phospholipid conjugates with cholesterol-phosphatidylcholine mixtures: sterically stabilized liposome formulations Bedu-Addo, F. K., P. Tang, et al. (1996), Pharm Res 13(5): 718-24. Abstract: PURPOSE. The purpose of this study was to investigate polyethyleneglycol (PEG)-phosphatidylethanolamine (PE) conjugate interaction with cholesterol-phospholipid mixtures in an attempt to explain the effect of cholesterol on liposome circulation time. METHODS. Differential scanning calorimetry, NMR, electron microscopy, dynamic light scattering and fluorescence spectroscopy were the major methods used. RESULTS. Studies performed in the absence of cholesterol indicated the formation of three distinct physical states depending on the chain length of PEG in PEG-PE. Mixed micelle formation was observed at concentrations of PEG(1,000)-DPPE above 7 mol-% of lipid. Phase separated lamellae were observed at all concentrations of PEG(12,000)-DPPE (Bedu-Addo et al. Pharm. Res. 13:710-717 (1996)). Upon incorporation of high concentrations of cholesterol >30 mol% into the lipid bilayer, the formation of phase separated lamellae was completely inhibited and the formation of mixed micelles significantly reduced. At high concentrations of PEG(1,000)-PE, solubilization of the bilayer occurred with preferential solubilization of cholesterol over phosphatidylcholine. Maximum steric stabilization (surface protection) was observed with low concentrations of short chain PEG-PE and high concentrations of cholesterol. CONCLUSIONS. The study provides a physical mechanism for the following observations: the blood circulation time is significantly increased or decreased with liposomes highly enriched with cholesterol or PEG-PE respectively. |
| Interaction of pulmonary surfactant protein A with dipalmitoylphosphatidylcholine and cholesterol at the air/water interface Yu, S. H. and F. Possmayer (1998), J Lipid Res 39(3): 555-68. Abstract: Interaction of pulmonary surfactant protein A (SP-A) with pure and binary mixed dipalmitoylphosphatidylcholine (DPPC) and cholesterol (3.5 wt%) at the air/saline, 1.5 mM CaCl2 interface was investigated using a rhomboid surface balance at 37 degrees C. Surface tension-area isotherms were measured to access the surface active properties of the monolayers. The organization of DPPC and cholesterol in DPPC and DPPC/cholesterol mixed monolayers with or without SP-A at equilibrium surface tension (approximately 23 mN/N) was revealed by autoradiographs of Langmuir-Blodgett (L-B) films deposited from 14CDPPC or 14Ccholesterol-labeled monolayers. The results showed that SP-A can interact with the polar head groups of DPPC monolayers and aggregate DPPC molecules. SP-A decreased the surface area reduction required for DPPC monolayers to achieve near zero surface tension from 30 to 25% of the area at equilibrium. SP-A also reduced the collapse surface tension of pure cholesterol from 27 to 23 mN/m. DPPC and cholesterol formed homogeneous mixed monolayers when both were dissolved in the spreading solvent prior to spreading, while separate cholesterol-rich domains appeared when DPPC and cholesterol were spread successively. Cholesterol resisted squeeze-out from either mixed monolayer through compression. Although SP-A could not promote the squeeze-out of cholesterol from homogeneous mixed monolayers, it facilitated that of cholesterol domains especially when SP-A had first interacted with DPPC. These results indicate that pulmonary surfactant protein A facilitates the squeeze-out of cholesterol domains from mixed monolayers by condensing DPPC and limiting lateral interactions of DPPC with cholesterol domains. |
| Interaction of reconstituted high density lipoprotein discs containing human apolipoprotein A-I (ApoA-I) variants with murine adipocytes and macrophages. Evidence for reduced cholesterol efflux promotion by apoA-I(Pro165-->Arg) von Eckardstein, A., G. Castro, et al. (1993), J Biol Chem 268(4): 2616-22. Abstract: Interaction of cells with both native and reconstituted high density lipoproteins (rHDL) containing apolipoprotein (apo) A-I mediates efflux of cellular cholesterol. To characterize structural requirements for this activity in apoA-I, six different genetic apoA-I variants and the corresponding normal allele products were isolated from heterozygous carriers, reconstituted with dimyristoylphosphatidylcholine (DMPC) into discoidal HDL and compared with regard to their ability to release intracellular cholesterol from murine adipocytes and peritoneal macrophages. In previous studies we determined the amino acid changes of these variants: Pro3-->Arg, Pro4-->Arg, Lys107-->0, Lys107-->Met, Pro165-->Arg, and Glu198-->Lys (von Eckardstein, A., Funke, H., Walter, M., Altland, K., Benninghoven, A., and Assmann, G. (1990) J. Biol. Chem. 265, 8610-8617) and demonstrated that all apoA-I variants except apoA-I(Lys107-->0) form discoidal HDL particles with phosphatidylcholine analogues indistinguishable from normal apoA-I (Jonas, A., von Eckardstein, A., Kezdy, K. E., Steinmetz, A., and Assmann, G. (1991) J. Lipid Res. 32, 95-106). In the present study, all apoA-I.DMPC complexes except those containing apoA-I(Pro165-->Arg) promoted cholesterol efflux as effectively as those containing normal apoA-I. Cholesterol efflux from adipocytes obtained after 180 min of incubation with apoA-I(Pro165-->Arg).DMPC complexes was decreased by 30% although this variant was bound to adipocytes with normal affinity. By contrast, apoA-I(Lys107-->Met).DMPC complexes were decreased in their binding affinity compared to normal apoA-I.DMPC complexes but normally promoted cholesterol efflux. Incubation of mouse peritoneal macrophages with apoA-I(Pro165-->Arg).DMPC complexes did also result in a 30% decreased efflux of radiolabeled cholesterol if compared to rHDL with the normal allele product from the same donor. Together the observations suggest that the substitution of proline at residue 165 interferes with the formation of a structural domain in apoA-I that is crucial for cellular cholesterol efflux stimulation. Furthermore, our results suggest that binding of HDL to adipocytes and cholesterol efflux promotion by HDL requires different structural domains. |
| Interaction of sex hormones and cholesterol with monolayers of dipalmitoylphosphatidylcholine in different phase state Dimitrov, O. A. and Z. I. Lalchev (1998), J Steroid Biochem Mol Biol 66(1-2): 55-61. Abstract: Interactions of estradiol, progesterone, testosterone and cholesterol with dipalmitoylphosphatidylcholine (DPPC) monolayers at the air/water interface using Wilhelmy-Tensiometer were studied by measuring the change of the monolayer surface tension gamma (mN/m). In order to estimate the role of DPPC phase state on the deltagamma effects the experiments were carried out at three temperatures: 37 degrees C, 41.5 degrees C and 47 degrees C, since at 37 degrees C and 47 degrees C the formation of ripple gel Pbeta and liquid-crystalline Lalpha DPPC phases respectively were realized. Surface tension lowering capacity of the individual components at the air/water interface decreases in the order cholesterol>estradiol>progesterone>testosterone. The surface tension decrease of previously formed DPPC monolayer after addition of cholesterol and hormones follows the order cholesterol>>estradiol>progesterone approximately = testosterone. The higher activity of cholesterol and estradiol is interpreted by the existence of hydroxyl group in the steroid A-ring and hydrocarbon chain in the cholesterol structure and the same hydroxyl group in the estradiol, with possible formation of hydrogen bond between this group and the C=O group of the DPPC. It is shown that the existence of C-H chain in the molecular structure is stronger determinant than the OH group regarding the interactions with the DPPC monolayers. The very low capacity of progesterone and testosterone to interact with the DPPC monolayer is explained by the lack of the C-H chain and OH group in their structures. It was shown that the interaction forces of the steroids studied with DPPC monolayers were dependent on the DPPC phase state, being in any conditions stronger in the Lalpha (47 degrees C) than in Pbeta (37 degrees C) phase. At 41.5 degrees C more complex behavior of the components at the monolayers was observed. The obtained results could serve the concept of regulated entry of the steroid sex hormones into the cells with the participation of the lipid membrane components. |