Liposome Methods and Protocols (Methods in Molecular Biology by Subhash C. Basu (Editor), Manju Basu (Editor)

By Subhash C. Basu (Editor), Manju Basu (Editor)

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Liposomes Composed of SM, Containing Domains of Cholesterol and of GM1 Ganglioside In these liposomes, distinct SM/cholesterol and SM/ganglioside domains coexist. The main characteristics of these liposomes are the following: size 100 nm (1000 Å); shape monolamellar; gel to liquid-crystalline temperature transition (Tm) 38°C, determined by high-sensitivity differential scanning Liposomes Containing Sphingolipids 23 calorimetry. Therefore, the liposomes are in the physical state of gel up to this temperature, and this feature should be taken into account anytime the physical state is important for the particular experiment.

4. 44 Pécheur and Hoekstra sample after gel filtration by Airfuge™ or SpeedVac™ (although this latter technique may lead to an undesired increase in the salt concentration, which may result in the swelling of liposomes when placed into a large volume of isotonic buffer). The WAE system or analogous systems employing different peptides offer the possibility to dissect at the molecular level the different steps of a fusion process. Indeed, aggregation between donor and target vesicles can be driven by the WAE peptide itself, via electrostatic interactions with a positively charged lipid incorporated in the target membrane, such as PE-lys (Fig.

129, 619–627. 9. , and Anderson, R. G. (1997) Tyrosine kinase receptors concentrated in caveolae-like domains from neuronal plasma membrane. J. Biol. Chem. 272, 3554–3559. 10. Parton, R. G. (1994) Ultrastructural localization of gangliosides; GM1 is concentrated in caveolae. J. Histochem. Cytochem. 42, 155–166. 11. Simons, K. and Van Meer, G. (1988) Lipid sorting in epithelial cells. Biochemistry 27, 6197–6202. 12. , and Freire, E. (1989) Influence of glycolipid oligosaccharide and long-chain base composition on the thermotropic properties of dipalmitoylphosphatidylcholine large unilamellar vesicles containing gangliosides Biochemistry 28, 5029–5039.

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