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• W2011276095 abstract "In the companion paper, I have reviewed the techniques employed for assessment of the asymmetric distribution and orientation of membrane proteins. This article deals with methods applicable to the investigation of the unequal distribution of lipids between the two membrane leaflets. Among the techniques I will discuss are the use of immunological techniques and lectins, chemical reagents, enzymatic isotopic labeling and degradation of membrane lipids, exchange proteins and physical techniques. Whenever appropriate, problems of crypticity and non-availability of lipids to interact with the appropriate ligands, reagents, modifying enzymes or exchange proteins have been envisaged. It appears that in many case, highly discordant results, sometimes with the same biological material, have been obtained. Some of the difficulties encountered presumably stem from the reported existence of non-bilayer arrangements and isotropic movement of lipids as evidenced by freeze-fracture and NMR studies. Other problems may be related to the induction of such arrangements, especially the inverted micellar arrangement, by the modifying agents, particularly degradation enzymes or exchange proteins when they cause severe unilateral modification of the lipids of the exposed leaflet. In addition, the situation is complicated by the role of the induced increase in the flip-flop rate under different experimental conditions and by modification of the rearrangement of lipid molecules as a result of the metabolic state of the cell or ghost preparation and of the reactivity of lipids as a consequence of temperature changes. Here, more so than with proteins, one must be cautious in interpreting experimental results. Moreover, it would appear that the use of different techniques in conjunction and the consequent comparison of results should be recommended. It has been emphasized that 'general rules' do not hold and that each new material should be assay again. To give one example, it is not pertinent to state that proteins enhance the flip-flop rate in lipid vesicles (and hence in membranes). This holds true for glycophorin from erythrocyte membrane, but could not be proved when mitochondrial cytochrome oxidase was used. There seems to be no rule for the distribution of lipids between the two leaflets of different membranes. For example, even for different strains of the same bacterial species, highly divergent results have been reported. It is generally (and probably under the influence of different studies with erythrocytes) believed that in mammalian plasma membranes, choline phospholipids are enriched in the outer leaflet and aminophospholipids in the inner leaflet. Though this contention may prove to be correct, different instances of contradictory results have been given in the text. This shows that if rules do exist, they remain to be discovered or established..." @default.
• W2011276095 created "2016-06-24" @default.
• W2011276095 creator A5032111035 @default.
• W2011276095 date "1980-12-01" @default.
• W2011276095 modified "2023-10-16" @default.
• W2011276095 title "Membrane asymmetry A survey and critical appraisal of the methodology II. Methods for assessing the unequal distribution of lipids" @default.
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