Membrane artificial lipid

Mechanisms of biological ion transport-carriers, channels and pumps in artificial lipid membranes. P. Lauger, Angew. Chem.,Int. Ed. Engl., 1985, 24, 905 (265). 

The ion channel receptors are relatively simple in functional terms because the primary response to receptor activation is generated by the ion channel which is an integral part of the protein. Therefore, no accessory proteins are needed to observe the response to nicotinic AChR activation and the full functioning of the receptor can be observed by isolating and purifying the protein biochemically and reconstituting the protein in an artificial lipid membrane. In contrast, the G-protein-coupled receptors require both G-proteins and those elements such as phospholipase-C illustrated in Fig. 3.1, in order to observe the response to receptor activation (in this case a rise in intracellular calcium concentration resulting from the action of IP3 on intracellular calcium stores). 

The lipid molecule is the main constituent of biological cell membranes. In aqueous solutions amphiphilic lipid molecules form self-assembled structures such as bilayer vesicles, inverse hexagonal and multi-lamellar patterns, and so on. Among these lipid assemblies, construction of the lipid bilayer on a solid substrate has long attracted much attention due to the many possibilities it presents for scientific and practical applications [4]. Use of an artificial lipid bilayer often gives insight into important aspects ofbiological cell membranes [5-7]. The wealth of functionality of this artificial structure is the result of its own chemical and physical properties, for example, two-dimensional fluidity, bio-compatibility, elasticity, and rich chemical composition. 

The survey of over 50 artificial lipid membrane models (pION) in this chapter reveals a new and very promising in vitro GIT model, based on the use of levels of lecithin membrane components higher than those previously reported, the use of negatively charged phospholipid membrane components, pH gradients, and artificial sink conditions. Also, a novel direction is suggested in the search for an ideal in vitro BBB model, based on the salient differences between the properties of the GIT and the BBB. 

A second approach with respect to anisotropic flavin (photo-)chemistry has been described by Trissl 18°) and Frehland and Trissl61). These authors anchored flavins in artificial lipid bilayers by means of C18-hydrocarbon chains at various positions of the chromophore. From fluorescence polarization analysis and model calculations they conclude, that the rotational relaxation time of the chromophore within the membrane is small compared to the fluorescence lifetime (about 2 ns74)). They further obtain the surprising result that the chromophore is localized within the water/lipid interface, with a tilt angle of about 30° (long axis of the chromophore against the normal of the membrane), irrespective of the position where the hydrocarbon chain is bound to the flavin nucleus. They estimate an upper limit of the microviscosity of the membrane of 1 Poise. 

At some point, these kinetic results for insulin on a biological membrane should be compared to kinetic results for insulin on an artificial lipid membrane, when such results become available. This comparison should be especially interesting in view of the suggestion by Sui et a/.(124) that nonspecific equilibrium binding of insulin to planar membranes is a function not only of membrane charge but also of some sort of nonelectrostatic mechanism, based on their TIRF experiments with a chamber adapted to a standard spectro-fluorimeter chamber. 

Of particular interest are membranes prepared of an inert porous support carrying natural or artificial lipids. These coatings may comprise a single component, such as isopropylmyristate or dodecanol [99, 106], or mixtures of comparable composition as the stratum corneum intercellular bilayer [107, 108], Usually, synthetic lipids are used, due to an elaborate isolation procedure for stratum corneum lipids, with limited yield and the necessity of separation of triglycerides, originating from subcutaneous fatty tissue or skin care products [109],... 

The possibility that pulmonary membranes are a primary site of ozone toxicity is suggested by a number of lines of evidence, most of which are indirect. These include observations that the membrane is the major site of ozone toxicity in plants and bacteria morphologic evidence of pulmonary membrane damage after ozone exposure in a number of studies and in vitro experiments with human red cells and artificial lipid membranes. 

Although artificial lipid membranes are almost impermeable to ions, biological membranes contain ion channels that selectively allow individual ion types to pass through (see p. 222). Whether an ion can cross this type of membrane, and if so in which direction, depends on the electrochemical gradient—i.e., on the concentrations of the ion on each side of the membrane (the concentration gradient) and on the difference in the electrical potential between the interior and exterior, the membrane potential. 

The model system for biological membranes is the so-called artificial lipid membrane made of two monolayers of lipid between two (aqueous) electrolyte solutions. The thickness of such a membrane can vary over a wide range and this introduces some ambiguity in deciding what kind of model should be the basis for the mathematical description of ion transport and other properties. Two extreme concepts can be distinguished. 

In the present study, therefore, lipid membranes were used as transducers of taste information. Artificial lipid materials, such as dioleyl phosphate (DOPH) or dioctadecyl-dimethyl-ammonium, were used to construct a lipid membrane and responses of electrical potential and resistance of the membranes were measured [9-15]. It was confirmed that the lipid membranes could discriminate five primary taste substances. Moreover, they could detect the interactions between taste substances observed in biological systems. The response properties were different in different types of lipids. If a hydrophobic part of a lipid was different, taste substances which can be detected were different. These facts indicate that the taste sensor can be realized by the use of various kinds of lipid membranes as transducers. 

A parallel development came from studies on artificial lipid bilayer membranes. Hladky and Hay don (1984) found that when very small amounts of the antibiotic gramicidin were introduced into such a membrane, its conductance to electrical current flow fluctuated in a stepwise fashion. It looked as though each gramicidin molecule contained an aqueous pore that would permit the flow of monovalent cations through it. Could the ion channels of natural cell membranes act in a similar way To answer this question, it was first necessary to solve the difficult technical problem of how to record the tiny currents that must pass through single channels. 

Artificial lipid membranes formed from phospholipid and steroids in both... 

The frequency response of various chemical constituents of nerve membrane was studied. Biological membranes in general consist of lipids and proteins. Firstly, impedance characteristics of artificial lipid bilayer membranes are examined using lecithin-hexadecane preparations. It was observed that the capacitance of plain lipid membranes was independent of frequency between 100 Hz and 20 KHz. Moreover, application of external voltages has no effect up to 200 mV. Secondly, membrane capacitance and conductance of nerve axon were investigated. There are three components in nerve membranes, i.e., conductance, capaci-... 

Channel activity is best studied electrochemically as charged species cross a cell membrane or artificial lipid bilayer. There is a difference in electrical potential between the interior and exterior of a cell leading to the membrane itself having a resting potential between -50 and -100 mV. This can be determined by placing a microelectrode inside the cell and measuring the potential difference between it and a reference electrode placed in the extracellular solution. Subsequent changes in electrical current or capacitance are indicative of a transmembrane flux of ions. 

Ions and small molecules may be transported across cell membranes or lipid bilayers by artificial methods that employ either a carrier or channel mechanism. The former mechanism is worthy of brief investigation as it has several ramifications in the design of selectivity filters in artificial transmembrane channels. To date there are few examples where transmembrane studies have been carried out on artificial transporters. The channel mechanism is much more amenable to analysis by traditional biological techniques, such as planar bilayer and patch clamp methods, so perhaps it is not surprising that more work has been done to model transmembrane channels. 

Clearly, major differences seem suggested in the mechanisms of hormone-membrane receptor interactions leading to various post-binding events [11], For FSH, as an example, there have been reports of hormone effects on membrane potentials of cultured Sertoli cells [12] and on conductance of artificial lipid membranes [13], as well as on amino acid transport systems [14]. Indeed, there has been one report of receptor-mediated gonadotropin action without receptor occupancy... 

For this purpose liposomes are used as lipid phase. Unilamellar liposomes are artificial lipid bilayer vesicles. They can be considered as real model bilayer membranes as they ideally consist of a circular bilayer membrane. The hydrophobic acyl chains are assembled in the hydrophobic core of the liposome whereas the hydrophilic head groups point to the water in the inside and outside of the vesicle. Liposomes can be produced from a variety of lipids and from mixtures of lipids. This possibility allows studying the influence of membrane constituents on the partition of solutes. Kramer et al. (1997) studied the influence of the presence of free fatty acids in membranes on the partition behaviour of propranolol. The influence on a-Tocopherol in membranes on the partition behaviour of desipramine has been reported recently (Marenchino et al. 2004) using a liposome model. 

The use of artificial lipid membranes and isolated membrane fragments and vesicles has been of great value in the study of membrane function, particularly transport and membrane-bound enzyme reactions. Current research in this area uses sophisticated techniques for determining the molec-... 

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