Phospholipid Vesicle Membranes Research Articles

Characteristics of phospholipid vesicles enhanced by adhesion on an annular region.

Phospholipid vesicle membranes are simple models used to study the mechanical properties of cell membranes. The shapes of flaccid vesicles can exhibit very diverse forms. When researching very flaccid vesicles, axisymmetrical vesicles with the membranes adhered to an annular region can also be observed. A phase diagram of such shapes was studied for different values of the vesicle parameters, i.e., the adhesion constant, the vesicle volume-to-membrane ratio, the volume ratio between the polar and the equatorial parts, and the equilibrium difference between the membrane monolayers. The energies of the annular shapes with respect to the vesicle parameters were closely examined and compared with the energies of the discocyte and stomatocyte shapes. The requirements for the existence of such annular shapes were also given for adhesion-free vesicle membranes. The results show that the adhesion between the lipid bilayers stabilizes the observed shapes, which belong to the locally stable branch of the annular vesicles. The value obtained for the adhesion constant of the SOPC membrane is 3×10^{-9}J/m^{2}.

Trans-membrane Fluorescence Enhancement by Carbon Dots: Ionic Interactions and Energy Transfer.

We report on trans-membrane interactions between blue-emitting carbon dots (CDs) and fluorescein. Hydrophobic CDs with a positive surface charge are embedded as-synthesized in the lipophilic sheet of the bilayer membrane of large synthetic phospholipid vesicles. The vesicles are prepared by mixing DOPC phospholipids and lipid molecules that contain anionic fluorescein attached to their hydrophilic head. Due to attractive electrostatic interactions, the CDs and fluorescein conjoin within the vesicle membrane, which leads to photoluminescence enhancement of fluorescein and facilitates trans-membrane energy transfer between the CDs and the dye.

A Fatty Acid Induces the Functional Assembly of a Channel Protein into Phospholipid Vesicles

The release of encapsulated cargo is an important step towards the design of intelligent drug delivery systems or vesicle-based bioreactors. Controlling the integration of pore forming channels into phospholipid vesicle membranes provides a route to initiate content release that could be coupled to gene-expression systems. Here, we present the design of phospholipid vesicles that release cargo upon the temporally controlled assembly of pore-forming channels. Our approach was to design a synthetic vesicle in which membrane composition could be remotely changed to enable the assembly of a pore-forming channel protein, α-hemolysin (αHL), and trigger content release. We first observed that a membrane lacking a sufficient concentration of cholesterol is not compatible with channel assembly and we hypothesized that increasing membrane fluidity, through the addition of oleic acid (OA) micelles could promote channel assembly. Using a dye release assay, we first confirmed that increasing OA content in phospholipid membranes leads to pore assembly. Next, we demonstrated that remote addition of OA micelles to inactive membranes results in fatty acid membrane uptake and channel assembly. We then showed that our system provides a route to control the integration of αHL into vesicle membranes when the channel protein is produced through a cell-free expression system. We remotely added OA micelles to vesicles present in a cell-free expression system where the amount of protein expressed is difficult to control. OA addition led to functional pore assembly and dye release. We ultimately demonstrate a new way to utilize remote changes in membrane composition to control αHL assembly and content release. Our study contributes to the design of vesicle-based technologies and also reveals new insight into the relationship between membrane composition and cholesterol-dependent cytolysin channel assembly.

Nitroalkylation of α-Synuclein by Nitro-Oleic Acid: Implications for Parkinson's Disease.

α-Synuclein (α-syn) represents the main component of the amyloid aggregates present in Parkinson's disease and other neurodegenerative disorders, collectively named synucleinopathies. Although α-syn is considered a natively unfolded protein, it shows great structural flexibility which allows the protein to adopt highly rich beta-sheet structures like protofibrils, oligomers and fibrils. In addition, this protein can adopt alpha-helix rich structures when interacts with fatty acids or acidic phospholipid vesicle membranes. When analyzing the toxicity of α-syn, protein oligomers are thought to be the main neurotoxic species by mechanisms that involve modification of intracellular calcium levels, mitochondrial and lysosomal function. Extracellular fibrillar α-syn promotes intracellular protein aggregation and shows many toxic effects as well. Nitro-fatty acids (nitroalkenes) represent novel pleiotropic anti-inflammatory signaling mediators that could interact with α-syn to exert unraveling actions. Herein, we demonstrated that nitro-oleic acid (NO2-OA) nitroalkylate α-syn, forming a covalent adduct at histidine-50. The nitroalkylated-α-syn exhibited strong affinity for phospholipid vesicles, moving the protein to the membrane compartment independent of composition of the membrane phospholipids. Moreover, NO2-OA-modified α-syn showed a reduced capacity to induce α-syn fibrillization compared to the non-nitrated oleic acid. From this data we hypothesize that nitroalkenes, in particular NO2-OA, may inhibit α-syn fibril formation exerting protective actions in Parkinson's disease.

Quenching of tryptophan fluorescence in a highly scattering solution: Insights on protein localization in a lung surfactant formulation.

CHF5633 (Chiesi Farmaceutici, Italy) is a synthetic surfactant developed for respiratory distress syndrome replacement therapy in pre-term newborn infants. CHF5633 contains two phospholipids (dipalmitoylphosphatidylcholine and 1-palmitoyl-2oleoyl-sn-glycero-3-phosphoglycerol sodium salt), and peptide analogues of surfactant protein C (SP-C analogue) and surfactant protein B (SP-B analogue). Both proteins are fundamental for an optimal surfactant activity in vivo and SP-B genetic deficiency causes lethal respiratory failure after birth. Fluorescence emission of the only tryptophan residue present in SP-B analogue (SP-C analogue has none) could in principle be exploited to probe SP-B analogue conformation, localization and interaction with other components of the pharmaceutical formulation. However, the high light scattering activity of the multi-lamellar vesicles suspension characterizing the pharmaceutical surfactant formulation represents a challenge for such studies. We show here that quenching of tryptophan fluorescence and Singular Value Decomposition analysis can be used to accurately calculate and subtract background scattering. The results indicate, with respect to Trp microenvironment, a conformationally homogeneous population of SP-B. Trp is highly accessible to the water phase, suggesting a surficial localization on the membrane of phospholipid vesicles, similarly to what observed for full length SP-B in natural lung surfactant, and supporting an analogous role in protein anchoring to the lipid phase.

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

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.

Pellet-free isolation of human and bovine milk extracellular vesicles by size-exclusion chromatography

ABSTRACTStudies have suggested that nanoscale extracellular vesicles (EV) in human and bovine milk carry immune modulatory properties which could provide beneficial health effects to infants. In order to assess the possible health effects of milk EV, it is essential to use isolates of high purity from other more abundant milk structures with well-documented bioactive properties. Furthermore, gentle isolation procedures are important for reducing the risk of generating vesicle artefacts, particularly when EV subpopulations are investigated. In this study, we present two isolation approaches accomplished in three steps based on size-exclusion chromatography (SEC) resulting in effective and reproducible EV isolation from raw milk. The approaches do not require any EV pelleting and can be applied to both human and bovine milk. We show that SEC effectively separates phospholipid membrane vesicles from the primary casein and whey protein components in two differently obtained casein reduced milk fractions, with one of the fractions obtained without the use of ultracentrifugation. Milk EV isolates were enriched in lactadherin, CD9, CD63 and CD81 compared to minimal levels of the EV-marker proteins in other relevant milk fractions such as milk fat globules. Nanoparticle tracking analysis and electron microscopy reveals the presence of heterogeneous sized vesicle structures in milk EV isolates. Lipid analysis by thin layer chromatography shows that EV isolates are devoid of triacylglycerides and presents a phospholipid profile differing from milk fat globules surrounded by epithelial cell plasma membrane. Moreover, the milk EV fractions are enriched in RNA with distinct and diverging profiles from milk fat globules. Collectively, our data supports that successful milk EV isolation can be accomplished in few steps without the use of ultracentrifugation, as the presented isolation approaches based on SEC effectively isolates EV in both human and bovine milk.

Fluorescent Probe Study of AOT Vesicle Membranes and Their Alteration upon Addition of Aniline or the Aniline Dimer p-Aminodiphenylamine (PADPA).

Artificial vesicles formed from sodium bis(2-ethylhexyl) sulfosuccinate (AOT) in aqueous solution are used successfully as additives for enzymatic oligomerizations or polymerizations of aniline or the aniline dimer p-aminodiphenylamine (PADPA) under slightly acidic conditions (e.g., pH 4.3 with horseradish peroxidase and hydrogen peroxide as oxidants). In these systems, the reactions occur membrane surface-confined. Therefore, (i) the physicochemical properties of the vesicle membrane and (ii) the interaction of aniline or PADPA with the AOT membrane play crucial roles in the progress and final outcome of the reactions. For this reason, the properties of AOT vesicles with and without added aniline or PADPA were investigated by using two fluorescent membrane probes: 1,6-diphenyl-1,3,5-hexatriene (DPH) and 6-lauroyl-2-dimethylaminonaphthalene (Laurdan). DPH and Laurdan were used as "sensors" of the membrane fluidity, surface polarity, and membrane phase state. Moreover, the effect of hexanol, alone or in combination with aniline or PADPA, as a possible modifier of the AOT membrane, was also studied with the aim of evaluating whether the membrane fluidity and surface polarity is altered significantly by hexanol, which, in turn, may have an influence on the mentioned types of reactions. The data obtained indicate that the AOT vesicle membrane at room temperature and pH 4.3 (0.1 M NaH2PO4) is more fluid and has a more polar surface than in the case of fluid phospholipid vesicle membranes formed from 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). Furthermore, the fluorescence measurements indicate that mixed AOT-hexanol membranes are less fluid than pure AOT membranes and that they have a lower surface polarity than pure AOT membranes. PADPA strongly binds to AOT and to mixed AOT/hexanol membranes and leads to drastic changes in the membrane properties (decrease in fluidity and surface polarity), resulting in Laurdan fluorescence spectra, which are characteristic for intramembrane phase separations (coexistence of ordered and disordered domains). This means that highly fluid AOT membranes transform upon the addition of PADPA into membranes that have ordered domains. Although the relevance of this finding for the enzymatic oligomerization of PADPA is not yet clear, it is also of interest if one likes to use heterogeneous vesicle membranes as additives for carrying out membrane surface-confined reactions that do not necessarily involve PADPA as a reactant.

Functionalized Vesicles with Co-Embedded CdSe Quantum Dots and [FeFe]-Hydrogenase Mimic for Light-Driven Hydrogen Production

A photocatalytic hybrid system for the production of hydrogen in purely aqueous solution was prepared by the co-embedding of hydrophobic oleic acid capped CdSe quantum dots (QD) for light harvesting and an amphiphilic [FeFe]-hydrogenase subunit mimic for proton reduction into phospholipid vesicle membranes. These bio-inspired functionalized vesicles concentrate the catalytic subunits in close proximity and show light-driven hydrogen generation in the presence of ascorbic acid resulting in 651 and 2.47 x 105 catalytic turnovers for the membrane-incorporated catalyst and the QDs, respectively.

Focus on Extracellular Vesicles: Development of Extracellular Vesicle-Based Therapeutic Systems.

Many types of cells release phospholipid membrane vesicles thought to play key roles in cell-cell communication, antigen presentation, and the spread of infectious agents. Extracellular vesicles (EVs) carry various proteins, messenger RNAs (mRNAs), and microRNAs (miRNAs), like a “message in a bottle” to cells in remote locations. The encapsulated molecules are protected from multiple types of degradative enzymes in body fluids, making EVs ideal for delivering drugs. This review presents an overview of the potential roles of EVs as natural drugs and novel drug-delivery systems.

Inward multivesiculation at the basal membrane of adherent giant phospholipid vesicles

Adherent giant vesicles composed of phosphatidylcholine, phosphatidylserine and biotinylated lipids form clusters of inward spherical buds at their basal membrane. The process is spontaneous and occurs when the vesicles undergo a sequence of osmotic swelling and deswelling. The daughter vesicles have a uniform size (diameter≈2–3μm), engulf small volumes of outer fluid and remain attached to the region of the membrane from which they generate, even after restoring the isotonicity. A pinning–sealing mechanism of long-wavelength modes of membrane fluctuations is proposed, by which the just-deflated vesicles reduce the surplus of membrane area and avoid excessive spreading and compression via biotin anchors. The work discusses the rationale behind the mechanism that furnishes GUVs with basal endovesicles, and its prospective use to simulate cellular events or to create molecular carriers.

Exosome-Mediated Targeted Delivery of miRNAs.

Many types of cells release phospholipid membrane vesicles that are thought to play key roles in cell-cell communication, antigen presentation, and the spread of infectious agents. These membrane vesicles, derived from the late endosomes, are called exosomes. Various proteins, messenger RNAs (mRNAs), and microRNAs (miRNAs) are carried by exosomes to cells in remote locations, like a message in a bottle. Because they can protect encapsulated small RNAs from ribonucleases (RNases) in body fluid, exosomes represent ideal carriers for nucleic acid drugs. In addition, because exosomes are constructed from self components, they are predicted to have low antigenicity and toxicity, extremely important properties for carriers used in drug delivery. This article describes a protocol for using exosomes as carriers for RNA drug delivery systems.

On the effect of serum on the transport of reactive oxygen species across phospholipid membranes.

The transport of plasma generated reactive oxygen species (ROS) across a simple phospholipid membrane mimic of a (real) cell was investigated. Experiments were performed in cell culture media (Dulbecco's modified Eagle's medium, DMEM), with and without 10% serum. A (broad spectrum) ROS reporter dye, 2,7-dichlorodihydrofluorescein (DCFH), was used to detect the generation of ROS by a helium (He) plasma jet in DMEM using free DCFH and with DCFH encapsulated inside phospholipid membrane vesicles dispersed in DMEM. The authors focus on the concentration and on the relative rates (arbitrary units) for oxidation of DCFH [or the appearance of the oxidized product 2,7-dichlorofluorescein (DCF)] both in solution and within vesicles. In the first 1 h following plasma exposure, the concentration of free DCF in DMEM was ~15× greater in the presence of serum (cf. to the serum-free DMEM control). The DCF in vesicles was ~2× greater in DMEM containing serum compared to the serum-free DMEM control. These data show that serum enhances plasma ROS generation in DMEM. As expected, the role of the phospholipid membrane was to reduce the rate of oxidation of the encapsulated DCFH (with and without serum). And the efficiency of ROS transport into vesicles was lower in DMEM containing serum (at 4% efficiency) when compared to serum-free DMEM (at 32% efficiency). After 1 h, the rate of DCFH oxidation was found to have significantly reduced. Based upon a synthesis of these data with results from the open literature, the authors speculate on how the components of biological fluid and cellular membranes might affect the kinetics of consumption of plasma generated ROS.

Helical peptaibol mimics are better ionophores when racemic than when enantiopure.

Helical peptide foldamers rich in α-aminoisobutyric acid (Aib) act as peptaibol-mimicking ionophores in the phospholipid bilayers of artificial vesicles. Racemic samples of these foldamers are more active than their enantiopure counterparts, which was attributed to differing propensities to form aggregates with crystal-like features in the bilayer.

1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC)-rich domain formation in binary phospholipid vesicle membranes: two-dimensional nucleation and growth.

Decades of study have probed phase transitions in model phospholipid bilayers and vesicles, especially in the context of the equilibrium phase diagram. Critical to the response of vesicles to environmental triggers, to the ultimate domain morphology, and to the approach to equilibrium (or not), we present here a study of domain formation in vesicles, focusing on a mechanism by which the cooling rate, tension, and composition affect the first appearance (nucleation) and subsequent growth of solid membrane domains. Employing a popular mixed membrane model based on DOPC and DPPC (1,2-dioleoyl-sn-glycero-3-phosphocholine and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine, respectively), we examined phase separation in giant two-component vesicles that were cooled from the one-phase fluid (Lα) region of the phase diagram into a region of fluid (Lα)-solid coexistence. At moderate and low membrane tensions, cooling produced solid DPPC-rich domains appearing as compact patches or irregular hexagons and likely with a Pβ' (ripple) arrangement. (The compact solid domains in this study differed distinctly from striped domains in vesicles of the same composition, in terms of molecular organization and conditions of first appearance during cooling.) The amounts of these solid domains were shown to adhere to the lever arm rule for a tie line on the phase diagram, with a solid composition near 95 mol % DPPC. The nucleation of the compact solid domains occurred in a short period, followed by rapid addition of ordered molecules to the nucleated domains, excluding tracer dye. The two-dimensional nucleation density of these compact solid domains (in the range of 10(-2)-10(-1) μm(-2)) was found to increase with the cooling rate (equivalent to the quench depth) with a greater than linear dependence. The 2-D nucleation density was also seen to decrease with membrane tension, presumably because membrane tension increases the line tension around a domain that opposes nucleation. A sigmoidal dependence of the nucleation density on the DPPC concentration was also found. With cooling rates in excess of ∼1 °C/min, solid domains persisted down to room temperature, likely passing from a preferred equilibrium to a local equilibrium with continued cooling. As a result of the persistence of the originally nucleated domains and the conservation of DPPC in the membrane, we observed an increasingly greater number of smaller domains with increased cooling rates. The domains in these vesicles were compact or hexagonal-shaped in contrast to flower-shaped dendritic domains in the same membrane system in a supported membrane configuration.

Confocal Raman Microscopy Probing of Temperature-Controlled Release from Individual, Optically-Trapped Phospholipid Vesicles

Control of permeability of phospholipid vesicle (liposome) membranes is critical to their applications in analytical sensing, in fundamental studies of chemistry in small volumes, and in encapsulation and release of payloads for site-directed drug delivery. Applications of liposome formulations in drug delivery often take advantage of the enhanced permeability of phospholipid membranes at their gel-to-fluid phase transition, where the release of encapsulated molecules can be initiated by an increase in temperature. Despite numerous successful liposome formulations for encapsulation and release methods to study the kinetics, this process has been limited to investigations of bulk vesicle dispersions, which provide little or no information about the vesicle membrane structure and its relationship to the kinetics of trans-membrane transport. In this work, confocal Raman microscopy is adapted to study temperature-dependent release of a model compound, 3-nitrobenzene sulfonate (3-NBS), from individual optically trapped phospholipid vesicles, while simultaneously monitoring structural changes in the vesicle membrane reported by vibrational modes of phospholipid acyl chains and the local environment of the encapsulated compound. The confocal geometry allows efficient excitation and collection of Raman scattering from a single vesicle, while optical trapping allows more than hour-long observations of the same vesicle. With window factor analysis to resolve component spectra, temperature-controlled release of 3-NBS through vesicle membranes composed of pure 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) was measured and compared to transport through a lysolipid-containing membrane specifically formulated for efficient drug delivery.

36 ENDOCYTOSIS UPTAKE OF LIPOSOMES IN UROTHELIUM CELLS DETECTED BY TRANSMISSION ELECTRON MICROSCOPY

INTRODUCTION AND OBJECTIVES: Intravesical liposomes (phospholipid membrane vesicles) have been used as a drug delivery platform and more recently, empty liposomes has shown promise as a treatment of interstitial cystitis/painful bladder syndrome. The exact mechanism for liposome uptake in bladder cells remains to be investigated and in the present study, we investigated the role of endocytosis in uptake of liposomes by urothelium cells. METHODS: Urothelial cell line (UROtsa) were left to adhere and grow for 48h in 100mm petridishes using Dulbecco modified Eagle medium (DMEM) containing 5% v/v fetal bovine serum with incubation at 37°C in a 5% CO2:95% air atmosphere. Liposomes encapsulating colloidal gold particles size 10nm were prepared by thin film hydration method and were incubated with serum free UROtsa cells for 2h either at 37°C or at 4°C. The morphology of liposome–cell interactions at two temperatures was assessed by transmission electron microscopy. RESULTS: Transmission Electron Microscopy (TEM) images located endocytic vesicles containing liposomes inside the UROtsa cells incubated at 37°C [panel B of attached picture] as indicated by dark stain of colloidal gold particles 10nm. In contrast, only extracellular binding was noticed in cells incubated at 4°C[panel A of attached picture]. Thin section electron micrographs show gold–containing liposomes trapped in intracellular vesicles, and in secondary lysosomes incubated at 37°C with colloidal gold liposomes. Absence of liposome internalization at 4°C indicate the need of energy dependent endocytosis as the primary mechanism of entry of liposomes into urothelium. CONCLUSIONS: Based on these observations, we propose that the main route of entry of liposomes into urothelium cells is by endocytosis and ultrastructural study provide evidence for a possible mechanism by which liposomes can act as drug delivery platform. These findings support the usefulness of liposomes in intravesical drug delivery. Source of Funding: NIDDK R01DK083323

The dynamics of melittin-induced membrane permeability

The transport of co-encapsulated solutes through the melittin-induced pores in the membrane of giant phospholipid vesicles was studied, and the characteristics of the pore formation process were modeled. Molecules of two different sizes (dextran and the smaller, fluorescent marker Alexa Fluor) were encapsulated inside the vesicles. The chosen individual vesicles were then transferred by micromanipulation from the stock suspension to the environment with the melittin (MLT). The vesicles were observed optically with a phase-contrast microscope and by monitoring the fluorescence signal. Such an experimental setup enabled an analysis of a single vesicle's response to the MLT on the basis of simultaneous, separate measurements of the outflow of both types of encapsulated molecules through the MLT-induced pores in the membrane. The mechanisms of the MLT's action were suggested in a model for MLT pore formation, with oligomeric pores continuously assembling and dissociating in the membrane. Based on the model, the results of the experiments were explained as a consequence of the membrane's permeability dynamics, with a continuously changing distribution of pores in the membrane with regard to their size and number. The relatively stable "average MLT pore" characteristics can be deduced from the proposed model.

Capacities of Membrane Lipids to Accumulate Neutral Organic Chemicals

Lipids have been considered as the predominant components for bioaccumulation of organic chemicals. However, differences in accumulation properties between different types of lipid (e.g., storage and membrane lipids) have rarely been considered. Moreover, in view of toxic effects on organisms, chemical accumulation specifically in biological membranes is of particular importance. In this review article, partition coefficients of 240 neutral organic compounds between liposomes (phospholipid membrane vesicles) and water (K(lipw)), reported in the literature or measured additionally for this work, were evaluated. Values of log K(lipw) and log K(ow) (octanol-water partition coefficients) differ by 0.4 on average. Polyparameter linear free energy relationships (PP-LFERs) can describe the log K(lipw) data even better (standard deviations = 0.28-0.31) than the log K(ow) model. Recent experimental data for highly hydrophobic compounds fit well to the PP-LFERs and do not indicate the existence of a previously postulated "hydrophobicity cutoff". Predictive approaches based only on the molecular structure (KOWWIN, SPARC, COSMOthermX, COSMOmic) were also evaluated for K(lipw) prediction. The PP-LFERs revealed that partition coefficients into membrane lipids can be two log units higher than those into storage lipids for H-bond donor compounds, suggesting that distinguishing between the two lipids is necessary to account for the bioaccumulation of these compounds, and that tissues rich in membrane lipids (e.g., kidneys, liver) instead of fat tissue can be the primary phase for accumulation.

Interaction of enterocyte FABPs with phospholipid membranes: Clues for specific physiological roles

Intestinal and liver fatty acid binding proteins (IFABP and LFABP, respectively) are cytosolic soluble proteins with the capacity to bind and transport hydrophobic ligands between different sub-cellular compartments. Their functions are still not clear but they are supposed to be involved in lipid trafficking and metabolism, cell growth, and regulation of several other processes, like cell differentiation. Here we investigated the interaction of these proteins with different models of phospholipid membrane vesicles in order to achieve further insight into their specificity within the enterocyte. A combination of biophysical and biochemical techniques allowed us to determine affinities of these proteins to membranes, the way phospholipid composition and vesicle size and curvature modulate such interaction, as well as the effect of protein binding on the integrity of the membrane structure. We demonstrate here that, besides their apparently opposite ligand transfer mechanisms, both LFABP and IFABP are able to interact with phospholipid membranes, but the factors that modulate such interactions are different for each protein, further implying different roles for IFABP and LFABP in the intracellular context. These results contribute to the proposed central role of intestinal FABPs in the lipid traffic within enterocytes as well as in the regulation of more complex cellular processes.