title: Asymmetric GUVs prepared by MβCD-mediated lipid exchange: an FCS study.
abstract: We report a simple method to obtain stable asymmetric giant unilamellar vesicles (GUVs). Fluorescence correlation spectroscopy was used to quantitatively characterize vesicle properties. After brain sphingomyelin (bSM) was exchanged into dioleoylphosphatidylcholine (DOPC) GUVs, lateral diffusion in the bSM-containing outer leaflet decreased, whereas that in the DOPC-containing inner leaflet was largely unchanged, confirming asymmetry and a lack of coupling between the physical states of the inner and outer leaflets. In contrast, after bSM was exchanged into brain phosphatidylcholine vesicles, lateral diffusion decreased in both leaflets. Thus, asymmetric GUVs should be useful for investigating the molecular mechanisms behind interleaflet coupling.
authors.affiliation: Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, New York, USA.
authors.forename: Salvatore
authors.initials: S
authors.lastname: Chiantia
authors.forename: Petra
authors.initials: P
authors.lastname: Schwille
authors.forename: Andrey S
authors.initials: AS
authors.lastname: Klymchenko
authors.forename: Erwin
authors.initials: E
authors.lastname: London
mesh_terms: D001921:Brain / Q000378:metabolism; D004058:Diffusion; D008055:Lipids / Q000737:chemistry*; D010713:Phosphatidylcholines / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013109:Sphingomyelins / Q000737:chemistry; D053835:Unilamellar Liposomes / Q000138:chemical synthesis*; D047392:beta-Cyclodextrins / Q000737:chemistry*
chemical_list: D008055:Lipids; D010713:Phosphatidylcholines; D013109:Sphingomyelins; D053835:Unilamellar Liposomes; D047392:beta-Cyclodextrins; C017251:1,2-oleoylphosphatidylcholine; C031215:betadex