Lipid phase states

An emergent field dealing with the physical/chemical processes that underlie the changes of lipid phase state during such cellular events as membrane fusion, vesicle trafficking, and cell disjunction. 

LIPID TRACER KINETICS Lipid phase state,... 

The experimental determination of the surface diffusion in foam films with FRAP technique is combined most successfully with the microinterferometric method of Scheludko-Exerowa (see Section 3.5) [39,40]. A systematic study showed the dependences of D on some film parameter such as film composition, type and film thickness, influence of proteins and lipid phase state, molecular length and charge, etc. (see Section 3.5.3). 

The dependence of D on lipid phase state was also reported [492], It was notable that the first measurable diffusion in the DMPC and DMPE thin films occurred at temperatures near the point at which the transition to the liquid crystalline La phase in bulk solution was observed. However, this is not the case for the negatively charged DMPG, for which diffusion (D = 610 8 cm2 s 1) was detected at two temperatures (15 and 20°C) where gel Plt and/or La phase may exist. 

There are many vital cellular processes, such as membrane fusion, which must involve, if only locally and transiently, changes in lipid phase state. To date, however, little attention has been paid to the dynamics of lipid phase interconversions. If, indeed, such changes are physiologically relevant, they must occur on a time scale comparable with those taking place in vivo. Thus, there is a need to establish the kinetics of lipid phase transitions. In addition to the kinetics, little is known about the mechanism of lipid phase transitions. Such information is integral to our understanding... 

A continuous lipidic cubic phase is obtained by mixing a long-chain lipid such as monoolein with a small amount of water. The result is a highly viscous state where the lipids are packed in curved continuous bilayers extending in three dimensions and which are interpenetrated by communicating aqueous channels. Crystallization of incorporated proteins starts inside the lipid phase and growth is achieved by lateral diffusion of the protein molecules to the nucleation sites. This system has recently been used to obtain three-dimensional crystals 20 x 20 x 8 pm in size of the membrane protein bacteriorhodopsin, which diffracted to 2 A resolution using a microfocus beam at the European Synchrotron Radiation Facility. 

Generally, the recrystaUization of S-layer protein into coherent monolayer on phospholipid films was demonstrated to depend on (1) the phase state of the hpid film, (2) the nature of the lipid head group (size, polarity, and charge), and (3) the ionic content and pH of the subphase [122,138] (Table 6). 

It has been shown by FM that the phase state of the lipid exerted a marked influence on S-layer protein crystallization [138]. When the l,2-dimyristoyl-OT-glycero-3-phospho-ethanolamine (DMPE) surface monolayer was in the phase-separated state between hquid-expanded and ordered, liquid-condensed phase, the S-layer protein of B. coagulans E38/vl was preferentially adsorbed at the boundary line between the two coexisting phases. The adsorption was dominated by hydrophobic and van der Waals interactions. The two-dimensional crystallization proceeded predominately underneath the liquid-condensed phase. Crystal growth was much slower under the liquid-expanded monolayer, and the entire interface was overgrown only after prolonged protein incubation. 

Figure 45-6. Interaction and synergism between antioxidant systems operating in the lipid phase (membranes) of the cell and the aqueous phase (cytosol). (R-,free radical PUFA-00-, peroxyl free radical of polyunsaturated fatty acid in membrane phospholipid PUFA-OOH, hydroperoxy polyunsaturated fatty acid in membrane phospholipid released as hydroperoxy free fatty acid into cytosol by the action of phospholipase Aj PUFA-OH, hydroxy polyunsaturated fatty acid TocOH, vitamin E (a-tocopherol) TocO, free radical of a-tocopherol Se, selenium GSH, reduced glutathione GS-SG, oxidized glutathione, which is returned to the reduced state after reaction with NADPH catalyzed by glutathione reductase PUFA-H, polyunsaturated fatty acid.)...

Most drug-like molecules dissolved in water form hydrogen bonds with the solvent. When such a molecule transfers from water into a phospholipid bilayer, the solute-water hydrogen bonds are broken (desolvation), as new solute-lipid H bonds form in the lipid phase. The free-energy difference between the two states of solvation has direct impact on the ability of the molecules to cross biological barriers. 

In order to learn about the phase states adopted by LPS and lipid A, Fourier-transform infrared spectroscopy, differential scanning calorimetry, and X-ray small-angle diffraction with CuXa or synchrotron radiation have been applied. In the following section, some recent results are summarized. 

Estimation or measurement of pKa is important to understand the state of ionization of the drug under physiological conditions and to evaluate salt-forming ability.27 Log P determines the partitioning of a drug between an aqueous phase and a lipid phase (i.e., lipid bilayer). Log P and acid pKa can be theoretically estimated with reasonable accuracy.14,28,29 High-throughput methods are also available for measurement of log P30 and pKa.31... 

Equation 7.1 utilizes exchange coefficients to predict steady-state BCFs and ATswS, and the model assumptions include a uniform lipid phase enclosed in a non-interactive membrane. The model shows that the magnitude of a BMO s BCF or an SPMD s Ksvj is affected by variations in ku and/or ke, unless both constants rise or fall proportionally. In the case of SPMDs, Huckins et al. (1993,2002a) have shown that the uptake and release process is essentially isotropic for HOCs. When residue exchange is isotropic, AfswS will remain relatively constant even when exposure conditions affect SPMD ku and ke values. This is not always the case for BMOs, yet isotropic exchange is a fundamental assumption of EP theory. 

The modulation of synaptosomal plasma membranes (SPMs) by adriamycin and the resultant effects on the activity of membrane-bound enzymes have been reported [58]. Again DPH was used as fluorescence probe. Adriamycin increased the lipid fluidity of the membrane labeled with DPH, as indicated by the steady-state fluorescence anisotropy. The lipid-phase separation of the membrane at 23.3 °C was perturbed by adriamycin so that the transition temperature was reduced to 16.2 °C. At the same time it was found that the Na+,K+-stimulated ATPase activity exhibits a break point at 22.8 °C in control SPMs. This was reduced to 15.8 °C in adriamydn-treated SPMs. It was proposed that adriamycin achieves this effect through asymmetric perturbation of the lipid membrane structure and that this change in the membrane fluidity may be an early key event in adriamycin-induced neurotoxicity. 

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