Reorganization of lipid domain distribution in giant unilamellar vesicles upon immobilization with different membrane tethers.
Characterization of phase coexistence in biologically relevant lipid mixtures is often carried out through confocal microscopy of giant unilamellar lipid vesicles (GUVs), loaded with fluorescent membrane probes. This last analysis is generally limited to the vesicle hemisphere further away from the coverslip, in order to avoid artifacts induced by the interaction with the solid surface, and immobilization of vesicles is in many cases required in order to carry out intensity, lifetime or single-molecule based microscopy. This is generally achieved through the use of membrane tethers adhering to a coverslip surface. Here, we aimed to determine whether GUV immobilization through membrane tethers induces changes in lipid domain distribution within liposomes displaying coexistence of lipid lamellar phases. Confocal imaging and a Förster resonance energy transfer (FRET) methodology showed that biotinylated phospholipids present significantly different membrane phase partition behavior upon protein binding, depending on the presence or absence of a linker between the lipid headgroup and the biotinyl moiety. Membrane phases enriched in a membrane tether displayed in some cases a dramatically increased affinity for the immobilization surface, effectively driving sorting of lipid domains to the adherent membrane area, and in some cases complete sequestering of a lipid phase to the interaction surface was observed. On the light of these results, we conclude that tethering of lipid membranes to protein surfaces has the potential to drastically reorganize the distribution of lipid domains, and this reorganization is solely dictated by the partition properties of the protein-tether complex.
Institute of Nanosciences and Nanotechnologies, Lisboa, Portugal.
M J
MJ
Sarmento
M
M
Prieto
Fábio
F
Fernandes
D001710:Biotin / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D008055:Lipids / Q000737:chemistry*; D008567:Membranes, Artificial; D008856:Microscopy, Fluorescence; D013050:Spectrometry, Fluorescence
D008055:Lipids; D008567:Membranes, Artificial; D001710:Biotin