Membrane lipid phase separation

Quinn, P.J. (1985). A lipid-phase separation model of low temperature damage to biological membranes. Cryobiology, 22, 128-40. 

As just mentioned, there are a large number of unsolved problems in membrane biophysics, including the questions of local anisotropic diffusion, hysteresis, protein-lipid phase separations, the role of fluctuations in membrane fusion, and the mathematical problems of diffusion in two dimensions Stokes paradox). 

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. 

Shaikh, S.R., et ah. Lipid phase separation in phospholipid bilayers and monolayers modeling the plasma membrane, Biochim. Biophys. Acta, 2001, 1512(2), 317-328. 

Membrane lipids exhibit complex polymorphism as a function of temperature. A balance of lipid phase structure is believed to result from interaction of lipids with other membrane components and solutes in the aqueous phase. In general, this balance results in a formation of a fluid lipid bilayer matrix. Phase separations of lipid from other membrane constituents can be driven by exposure of membranes to temperatures outside the normal growth temperature. These can be the creation of gel phase domains at low temperature or the formation of nonbilayer structures at high temperature. Both types of lipid phase separation are associated with functional changes in the membrane including loss of selective permeability barrier properties. 

Non-ionic surfactants (PEG esters of C2-C18 fatty acids) have been found to modulate polymorphonuclear leukocyte locomotion [164]. Esters with shorter aliphatic chains had negligible effect whereas those with longer hydrocarbon chains (C16, Cig) reduced locomotion, an effect perhaps mediated by alteration of membrane structure, for instance inducing lateral lipid phase separation which might alter the anchoring of the cytoskeleton and the position of membrane constituents responsible for adhesion of the cells to other cells and to various substrates [165]. 

Table 1. Midpoint values for the critical temperatures of lipid phase separation of cytoplasmic membrane, R -release, and rate of photosynthetic oxygen evolution tested in control and hydrogenated A. nidulans cells.

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)... 

Considering only the lipid phase as the transport pathway for the peptide, as the solute enters and diffuses across the membrane it will encounter a number of different microenvironments. The first is the aqueous membrane interface (Fig. 23). In this region, the hydrated polar headgroups of the membrane phospholipids separate the aqueous phase from the apolar membrane interior. It has been shown that this region is capable of satisfying up to 70% of the hydrophobic effect... 

Recently, due to increased interest in membrane raft domains, extensive attention has been paid to the cholesterol-dependent liquid-ordered phase in the membrane (Subczynski and Kusumi 2003). The pulse EPR spin-labeling DOT method detected two coexisting phases in the DMPC/cholesterol membranes the liquid-ordered and the liquid-disordered domains above the phase-transition temperature (Subczynski et al. 2007b). However, using the same method for DMPC/lutein (zeaxanthin) membranes, only the liquid-ordered-like phase was detected above the phase-transition temperature (Widomska, Wisniewska, and Subczynski, unpublished data). No significant differences were found in the effects of lutein and zeaxanthin on the lateral organization of lipid bilayer membranes. We can conclude that lutein and zeaxanthin—macular xanthophylls that parallel cholesterol in its function as a regulator of both membrane fluidity and hydrophobicity—cannot parallel the ability of cholesterol to induce liquid-ordered-disordered phase separation. 

The properties of membranes commonly studied by fluorescence techniques include motional, structural, and organizational aspects. Motional aspects include the rate of motion of fatty acyl chains, the head-group region of the phospholipids, and other lipid components and membrane proteins. The structural aspects of membranes would cover the orientational aspects of the lipid components. Organizational aspects include the distribution of lipids both laterally, in the plane of the membrane (e.g., phase separations), and across the membrane bilayer (phospholipid asymmetry) and distances from the surface or depth in the bilayer. Finally, there are properties of membranes pertaining to the surface such as the surface charge and dielectric properties. Fluorescence techniques have been widely used in the studies of membranes mainly since the time scale of the fluorescence lifetime coincides with the time scale of interest for lipid motion and since there are a wide number of fluorescence probes available which can be used to yield very specific information on membrane properties. 

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. 

Membrane conformational changes are observed on exposure to anesthetics, further supporting the importance of physical interactions that lead to perturbation of membrane macromolecules. For example, exposure of membranes to clinically relevant concentrations of anesthetics causes membranes to expand beyond a critical volume (critical volume hypothesis) associated with normal cellular function. Additionally, membrane structure becomes disorganized, so that the insertion of anesthetic molecules into the lipid membrane causes an increase in the mobility of the fatty acid chains in the phospholipid bilayer (membrane fluidization theory) or prevent the interconversion of membrane lipids from a gel to a liquid form, a process that is assumed necessary for normal neuronal function (lateral phase separation hypothesis). 

Ringsdorf, Sackmann, and coworkers characterized the behavior of mixtures of the polymerizable bis-dienoylammonium lipid 14 and DMPC [42]. Evidence for phase separation in these mixtures was obtained from electron microscopy and light scattering. Since the intensity of scattered light is dependent on the physical state of the membrane, plots of scattering intensity versus temperature exhibit inflections at phase transitions. This technique was used in conjunction... 

---