Evolution of lipidic structures during model membrane fusion and the relation of this process to cell membrane fusion

Biochemistry
J Lee, B R Lentz

Abstract

The sequence of events involved in poly(ethylene glycol)-mediated fusion of small unilamellar vesicles (SUVs) has been studied. Fusion events were monitored using light scattering for vesicle aggregation, the fluorescence lifetime of membrane probe lipids (DPHpPC and NBD-PS) for membrane mixing, the aqueous fluorescent marker (Tb3+/DPA and H+/HPTS) for contents mixing; and quasi-elastic light scattering for the change in the size of vesicles. Poly(ethylene glycol) is a highly hydrated polymer that can bring vesicle membranes to near molecular contact but is unable to induce vesicle fusion without manipulations that reduce packing density and encourage molecular motions in the backbone regions of both contacting membrane leaflets. Once this condition is achieved, the sequence of events involved in vesicle fusion is shown here to be (1) outer leaflet mixing accompanied by (2) transient pore formation, both occurring on a time scale of approximately 10 s and leading to an initial, reversible intermediate; (3) a 1-3 min delay leading to formation of a fusion-committed second intermediate; (4) inner leaflet mixing on a time scale of ca. 150 s; and (5) contents mixing on a time scale of 150-300 s. Inner leaflet mixing, which has neve...Continue Reading

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