Osmotic and curvature stress affect PEG-induced fusion of lipid vesicles but not mixing of their lipids.
Poly (ethylene glycol) (PEG) in the external environment of membrane vesicles creates osmotic imbalance that leads to mechanical stress in membranes and may induce local membrane curvature. To determine the relative importance of membrane stress and curvature in promoting fusion, we monitored contents mixing (CM) and lipid mixing (LM) between different sized vesicles under a variety of osmotic conditions. CM between highly curved vesicles (SUV, 26 nm diameter) was up to 10 times greater than between less curved vesicles (LUV, 120 nm diameter) after 5 min incubation at a low PEG concentration (<10 wt%), whereas LM was only approximately 30% higher. Cryo-electron microscopy showed that PEG at 10 wt% did not create high curvature contacts between membranes in LUV aggregates. A negative osmotic gradient (-300 mOs/kg, hypotonic inside) increased CM two- to threefold for both types of vesicles, but did not affect LM. A positive gradient (+220 mOs/kg, hypertonic inside) nearly eliminated CM and had no effect on LM. Hexadecane added to vesicles had no effect on LM but enhanced CM and reduced the inhibitory effect on CM of a positive osmotic gradient, but had little influence on results obtained under a negative osmotic gradient. We conclude that the ability of closely juxtaposed bilayers to form an initial intermediate ("stalk") as soon as they come into close contact was not influenced by osmotic stress or membrane curvature, although pore formation was critically dependent on these stresses. The results also suggest that hexadecane affects the same part of the fusion process as osmotic stress. We interpret this result to suggest that both a negative osmotic gradient and hexadecane reduce the unfavorable free energy of hydrophobic interstices associated with the intermediates of the fusion process.
Malinin
Vladimir S
VS
Department of Biochemistry and Biophysics, CB#7260, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7260, USA.
Frederik
Peter
P
Lentz
Barry R
BR
D000473:Alkanes / Q000737:chemistry; D055592:Biophysical Phenomena; D001703:Biophysics; D020285:Cryoelectron Microscopy; D004305:Dose-Response Relationship, Drug; D008055:Lipids / Q000737:chemistry*; D015233:Models, Statistical; D009994:Osmolar Concentration; D009995:Osmosis; D011092:Polyethylene Glycols / Q000737:chemistry*
D000473:Alkanes; D008055:Lipids; D011092:Polyethylene Glycols; C007932:n-hexadecane