Model membranes, phospholipids

A very brief description of biological membrane models, and model membranes, is given. Studies of lateral diffusion in model membranes (phospholipid bilayers) and biological membranes are described, emphasizing magnetic resonance methods. The relationship of the rates of lateral diffusion to lipid phase equilibria is discussed. Experiments are reported in which a membrane-dependent immunochemical reaction, complement fixation, is shown to depend on the rates of diffusion of membrane-bound molecules. It is pointed out that the lateral mobilities and distributions of membrane-bound molecules may be important for cell surface recognition. 

When placed in aqueous solution, phospholipids spontaneously form lipid bilayers. According to the fluid-mosaic model, membrane phospholipids form lipid bilayers with membrane proteins associated with the bilayer as both peripheral and integral proteins. 

In 1972, S. J. Singer and G. L. Nicolson proposed the fluid mosaic model for membrane structure, which suggested that membranes are dynamic structures composed of proteins and phospholipids. In this model, the phospholipid bilayer is a fluid matrix, in essence, a two-dimensional solvent for proteins. Both lipids and proteins are capable of rotational and lateral movement. 

Papahadjopoulos, D., and Watkins, J. C. (1967). Phospholipid model membranes. II. Permeability properties of hydrated liquid crystals, Biochim. Biophys. Acta. 135. 639-652. 

FIG. 14 Schematic illustration of an archaeal cell envelope structure (a) composed of the cytoplasmic membrane with associated and integral membrane proteins and an S-layer lattice, integrated into the cytoplasmic membrane, (b) Using this supramolecular construction principle, biomimetic membranes can be generated. The cytoplasmic membrane is replaced by a phospholipid or tetraether hpid monolayer, and bacterial S-layer proteins are crystallized to form a coherent lattice on the lipid film. Subsequently, integral model membrane proteins can be reconstituted in the composite S-layer-supported lipid membrane. (Modified from Ref. 124.)... 

Different tissues have different lipid compositions. The most common lipid components of membranes are PC and PE. Lipid extracts from brain and lung are also rich in PS heart tissue is rich in PG, and liver is rich in PI [567]. Human blood cells, as ghost erythrocytes (with cytoplasm contents removed), are often used as membrane models. These have different compositions between the inner and outer leaflets of the bilayer membrane. Phospholipids account for 46% of the outer leaflet membrane constituents, with PC and Sph about equal in amount. The inner leaflet is richer in phospholipids (55%), with the mix 19% PE, 12% PS, 7% PC and 5% Sph [567],... 

Rachel, K., Asuncionpunzalan, E. and London, E. (1995) Anchoring of tryptophan and tyrosine analogs at the hydrocarbon polar boundary in model membrane-vesicles - paralax analysis of fluorescence quenching induced by nitroxide-labelled phospholipids. Biochemistry 34,15475-15479. 

Recently in our group, model membrane permeation barriers have been constructed with concentrated phospholipid solutions, 10-74% wt/vol soy lecithin (approximate %w/w lipid composition 24% PC, 18% PE, 12% PI cf. Table 3.1) in dodecane, supported on high-porosity, hydrophobic microfilters. This newly formulated lipid has a net negative charge at pH 7.4, which further increases above pH 8, as the ethanolamine groups deionize. Also tested were 10% wt/vol egg lecithin lipid solutions in dodecane (approximate composition 73% PC, 11% PE,... 

Yasuda, T., Dancey, G.F., and Kinsky, S.C. (1977) Immunogenicity of liposomal model membranes in mice Dependence on phospholipid composition. Proc. Natl. Acad. Sci. USA 74, 1234-1236. 

Abstract To understand how membrane-active peptides (MAPs) function in vivo, it is essential to obtain structural information about them in their membrane-bound state. Most biophysical approaches rely on the use of bilayers prepared from synthetic phospholipids, i.e. artificial model membranes. A particularly successful structural method is solid-state NMR, which makes use of macroscopically oriented lipid bilayers to study selectively isotope-labelled peptides. Native biomembranes, however, have a far more complex lipid composition and a significant non-lipidic content (protein and carbohydrate). Model membranes, therefore, are not really adequate to address questions concerning for example the selectivity of these membranolytic peptides against prokaryotic vs eukaryotic cells, their varying activities against different bacterial strains, or other related biological issues. 

Prosser RS, Hwang JS, Void RR (1998) Magnetically aligned phospholipid bilayers with positive ordering a new model membrane system. Biophys J 74 2405-2418... 

Conformational changes in BmorPBP were also studied in the presence of model membranes using CD spectroscopy. Conformational changes more pronounced than those observed at low pH were detected in the presence of anionic vesicles of dimyristoylphosphatidylglycerol (DMPG), whereas the effect of neutral phospholipids vesicles, dimyristoylphosphatidylcholine (DMPC) was... 

E. London and G. W. Feigenson, Fluorescence quenching in model membranes. 1. Characterization of quenching caused by a spin-labeled phospholipid, Biochemistry 20, 1932-1938 (1981). 

Natural biological membranes consist of lipid bilayers, which typically comprise a complex mixture of phospholipids and sterol, along with embedded or surface associated proteins. The sterol cholesterol is an important component of animal cell membranes, which may consist of up to 50 mol% cholesterol. As cholesterol can significantly modify the bilayer physical properties, such as acyl-chain orientational order, model membranes containing cholesterol have been studied extensively. Spectroscopic and diffraction experiments reveal that cholesterol in a lipid-crystalline bilayer increases the orientational order of the lipid acyl-chains without substantially restricting the mobility of the lipid molecules. Cholesterol thickens a liquid-crystalline bilayer and increases the packing density of lipid acyl-chains in the plane of the bilayer in a way that has been referred to as a condensing effect. 

Lateral diffusion of phospholipids in model membranes at ambient pressure has been studied over the years by a variety of techniques including fluorescence recovery after photobleaching (FRAP), spin-label ESR, pulse field gradient NMR (PFG-NMR), quasielastic neutron scattering (QENS), excimer fluorescence and others.In general, the values reported for the lateral diffusion coefficient (D) range from 10 to 10 cm /s in the... 

Fig. 4. Schematic representation of transient method employed by Devaux and McConnell9 to measure the rates of lateral diffusion of phospholipids in model membranes. The upper diagram represents a concentrated patch of labels at the beginning of the experiment, time f = 0. At later times f>0, the molecules diffuse laterally, as shown in the lower two drawings. The paramagnetic resonance spectra depend on the spin-label concentration in the plane of the membrane, and an analysis of the time dependence of these spectra yielded the diffusion constant. [Reprinted with permission from P. Devaux and H. M. McConnell, J. Am. Chem. Soc., 94, 4475 (1972). Copyright by American Chemical Society.]...

In a very thoughtful investigation of solvent systems to model membrane characteristics, Leahy et al. (1989, 1992) have argued that two receptors sited in different tissues (or membranes) could exist in environments that are very different in hydrogen bonding character one may be surrounded by amphiprotic groups (as in a protein) or by proton donors the other may be surrounded by proton acceptors (as in a phospholipid membrane). 

Qeveral recent investigations using various physicochemical methods have provided convincing evidence to support the contention that the basic structure of most biological membranes consists of a phospholipid bilayer (1,2,3, 4). Studies on phospholipid model membranes can therefore be expected to yield relevant information on the role played by phospholipids in determining the characteristic properties of biological membranes (5). One important aspect of this problem concerns the mechanisms of interaction between the phospholipids and other membrane constituents such as cholesterol, proteins, and different inorganic... 

Fig. 4.4 Hypothetical model showing the modulation of glutamate transporter by arachidonic acid. Interactions of glutamate with its receptor result in depolarization and Ca2+ entry into the cell. Ca2+-mediated stimulation of PLA2 results in breakdown of neural membrane phospholipids and the release of arachidonic acid. Arachidonic acid not only modulates proton conductance associated with neuronal excitability, but also provides eicosanoids, which may control the glutamate transporter (modified from Fairman and Amara, 1999)...

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