Membrane lipid bilayers model systems

Lipids in model systems are often found in asymmetric clusters (see Figure 9.8). Such behavior is referred to as a phase separation, which arises either spontaneously or as the result of some extraneous influence. Phase separations can be induced in model membranes by divalent cations, which interact with negatively charged moieties on the surface of the bilayer. For example, Ca induces phase separations in membranes formed from phosphatidylserine (PS)... 

Hydrated bilayers containing one or more lipid components are commonly employed as models for biological membranes. These model systems exhibit a multiplicity of structural phases that are not observed in biological membranes. In the state that is analogous to fluid biological membranes, the liquid crystal or La bilayer phase present above the main bilayer phase transition temperature, Ta, the lipid hydrocarbon chains are conforma-tionally disordered and fluid ( melted ), and the lipids diffuse in the plane of the bilayer. At temperatures well below Ta, hydrated bilayers exist in the gel, or Lp, state in which the mostly all-trans chains are collectively tilted and pack in a regular two-dimensional... 

This review addresses the issues of the chemical and physical processes whereby inorganic anions and cations are selectively retained by or passed through cell membranes. The channel and carrier mechanisms of membranes permeation are treated by means of model systems. The models are the planar lipid bilayer for the cell membrane, Gramicidin for the channel mechanism, and Valinomycin for the carrier mechanism. 

With the adequacy of lipid bilayer membranes as models for the basic structural motif and hence for the ion transport barrier of biological membranes, studies of channel and carrier ion transport mechanisms across such membranes become of central relevance to transport across cell membranes. The fundamental principles derived from these studies, however, have generality beyond the specific model systems. As noted above and as will be treated below, it is found that selective transport... 

A final example of the simulation of a complex system is a series of MD simulations of bilayer membranes. Membranes are crucial constituents of living organisms they are the scene for many important biological processes. Experimental data are known for model systems for example for the system sodium decanoate, decanol and water that forms smectic liquid crystalline structures at room temperature, with the lipids organized in bilayers. 

While the fluid mosaic model of membrane stmcture has stood up well to detailed scrutiny, additional features of membrane structure and function are constantly emerging. Two structures of particular current interest, located in surface membranes, are tipid rafts and caveolae. The former are dynamic areas of the exo-plasmic leaflet of the lipid bilayer enriched in cholesterol and sphingolipids they are involved in signal transduction and possibly other processes. Caveolae may derive from lipid rafts. Many if not all of them contain the protein caveolin-1, which may be involved in their formation from rafts. Caveolae are observable by electron microscopy as flask-shaped indentations of the cell membrane. Proteins detected in caveolae include various components of the signal-transduction system (eg, the insutin receptor and some G proteins), the folate receptor, and endothetial nitric oxide synthase (eNOS). Caveolae and lipid rafts are active areas of research, and ideas concerning them and their possible roles in various diseases are rapidly evolving. 

Here, we discuss a solid-state 19F-NMR approach that has been developed for structural studies of MAPs in lipid bilayers, and how this can be translated to measurements in native biomembranes. We review the essentials of the methodology and discuss key objectives in the practice of 19F-labelling of peptides. Furthermore, the preparation of macroscopically oriented biomembranes on solid supports is discussed in the context of other membrane models. Two native biomembrane systems are presented as examples human erythrocyte ghosts as representatives of eukaryotic cell membranes, and protoplasts from Micrococcus luteus as membranes... 

Because membranes components participate in nearly every cell activity their structures are also dynamic and far from the equilibrium states that are most readily understood in biophysical terms. Newly synthesized bilayer lipids are initially associated with endoplasmic reticulum (Ch.3) whereas phospholipids initially insert into the cytoplasmic leaflet while cholesterol and sphingolipids insert into the luminal endoplasmic reticulum (ER) leaflet. Glycosylation of ceramides occurs as they transit the Golgi compartments, forming cerebrosides and gangliosides in the luminal leaflet. Thus, unlike model systems, the leaflets of ER membranes are asymmetric by virtue of their mode of biosynthesis. 

The studies on phospholipid bilayers with defined amounts of charged component are helpful to explain the partition characteristics in biological membranes. Liposome water partition data of propranolol in lipids from kidney epithelial cells (a common model system in pharmaceutical sciences for the uptake into the gastrointestinal tract) have been successfully described with partition models developed for pure bilayers or defined mixtures [159]. Since lipophilic cations and anions can be used as probes for the membrane potential, their interaction with microbial and mitochondrial membranes has been studied... 

The use of DPH lifetimes for the analysis of phase separations and membrane properties has been described for mode) systems.n fl) In the case of both parinaric acids and DPH, one of the motivations for examining phase separation in a model lipid bilayer is the possibility that phase separations might be detectable in natural membranes. However, this technique has not been able to satisfactorily resolve lateral phase separations in natural membranes, either because they do not exist or because they are much more complex and even possibly transient in nature. Alternatively, it could be argued that if a probe could be found with the characteristics of trans-parinaric acid but perhaps with an even greater phase partitioning ability, then this approach might be reevaluated. 

L. Davenport, R. E. Dale, R. H. Bisby, and R. B. Cundall, Transverse location of the fluorescent probe l,6-diphenyl-l,3,5-hexatriene in model lipid bilayer membrane systems by resonance excitation energy transfer, Biochemistry 24, 4097-4108 (1985). 

This permeability barrier shows selectivity in that small hydrophobic molecules can partition into and diffuse across the lipid bilayer of the cell membrane, whereas small hydrophilic molecules can only diffuse between cells (i.e., through the intercellular junctions). In addition, the presence of uptake and efflux transporters complicates our ability to predict intestinal permeability based on physicochemical properties alone because transporters may increase or decrease absorptive flux. The complexity of the permeability process makes it difficult to elucidate permeability pathways in complex biological model systems such as animals and tissues. For this reason, cultured cells in general, and Caco-2 cells in particular, have been used extensively to investigate the role of specific permeability pathways in drug absorption. 

An important approach to the study of biological membranes has been the preparation and study of model membranes. According to current usage, model membranes include lipid bilayers and lipid bilayers into which have been incorporated additional components such as one or more membrane proteins. It is through the study of such model membranes that one has the best opportunity to isolate and study fundamental physical chemical and biophysical processes, and it is for this reason that the present report emphasizes these systems. A discussion of model membranes necessarily starts with a description of the chemical compositions and physical properties of lipid molecules. 

At the most fundamental level, monolayers of surfactants at an air-liquid interface serve as model systems to examine condensed matter phenomena. As we see briefly in Section 7.4, a rich variety of phases and structures occurs in such films, and phenomena such as nucleation, dendritic growth, and crystallization can be studied by a number of methods. Moreover, monolayers and bilayers of lipids can be used to model biological membranes and to produce vesicles and liposomes for potential applications in artificial blood substitutes and drug delivery systems (see, for example, Vignette 1.3 on liposomes in Chapter 1). 

Much of our understanding of the chemical aspects of cell membranes has been derived from model systems based on surfactants, especially membrane lipids. In this section we are primarily concerned with the use of monolayers, bilayers, and especially black lipid membranes and vesicles as cell membrane models. 

The discriminative uptake of alkali metal cations by biological systems, through their membranes, has been an area of much interest. In the membrane, the cations must pass through a lipid bilayer of low dielectric constant and this has led to the proposition that the cation could be selectively transferred via a carrier molecule, or through a suitably donor-lined pore.7-9 As a consequence of their selective properties, the polyethers and cryptands have been investigated as speculative models for the above process and selectivity sequences have been established. 

Another approach to a combined system is the connection of the two systems through a quinone redox couple dissolved in an oil phase, as shown in Fig. 17.2. This system is analogous to the combination of photosystems I and II in the photosynthesis of green plants. Fig. 17.10 illustrates the structure of our model system, in which the oil-phase corresponds to the lipid bilayer membrane of chloroplast. Such a system is structurally identical to a liposome and has the possibility of development for use in a batch reactor. 

Conventional liposomes and lipid complexes. Liposomes were used initially as a model system for cellular membranes to study the biochemistry of membrane proteins.85 Consequently, when liposomes were first tried as a drug delivery system, their bilayers were composed of un-derivatized naturally occurring lipids. Most of such conventional liposomes are taken up by the MPS phagocytes within a few hours of injection, mostly by liver Kupffer cells and spleen macrophages.9 Inside the endosomes and lysosomes of those cells, liposomes are degraded. If the liposomal drugs are membrane permeable, they then can diffuse from the endosomal compartments to the cytoplasm of the macrophage cells and slowly reenter the blood circulation. Because such a clearance... 

The newly synthesized phenothiazine derivative 2-trifluoromethyl-10-[4-(methanesulfonylamido)butyl]-phenothiazine (FPhMS, see Fig. 1 for chemical structure) was also extensively studied in the context of its interaction with lipid bilayers. DSC was used to study the influence of this compound on model membranes formed from DMPE [80], DPPC [81], DMPC, and DMPG [82]. In all the studied lipid systems FPhMS (16) lowered Tm, caused broadening of transition peaks, and induced the decrease of AH. Melting temperatures were found to be reduced by the phenothiazine derivative to a similar extent when different lipids possessing acyl chains of the same length were compared. 

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