Influence of phospholipid asymmetry on fusion between large unilamellar vesicles.
The ability of lipid asymmetry to regulate Ca(2+)-stimulated fusion between large unilamellar vesicles has been investigated. It is shown that for 100-nm-diameter LUVs composed of dioleoylphosphatidylcholine, dioleoylphosphatidylethanolamine, phosphatidylinositol, and dioleoylphosphatidic acid (DOPC/DOPE/PI/DOPA; 25:60:5:10) rapid and essentially complete fusion is observed by fluorescent resonance energy transfer techniques when Ca2+ (8 mM) is added. Alternatively, for LUVs with the same lipid composition but when DOPA was sequestered to the inner monolayer by incubation in the presence of a pH gradient (interior basic), little or no fusion is observed on addition of Ca2+. It is shown that the extent of Ca(2+)-induced fusion correlates with the amount of exterior DOPA. Further, it is shown that LUVs containing only 2.5 mol % DOPA, but where all the DOPA is in the outer monolayer, can be induced to fuse to the same extent and with the same rate as LUVs containing 5 mol % DOPA. These results strongly support a regulatory role for lipid asymmetry in membrane fusion and indicate that the fusogenic tendencies of lipid bilayers are largely determined by the properties of the monolayers proximate to the fusion interface.
Eastman
S J
SJ
Department of Biochemistry, University of British Columbia, Vancouver, Canada.
Hope
M J
MJ
Wong
K F
KF
Cullis
P R
PR
D002118:Calcium; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D005614:Freeze Fracturing; D006863:Hydrogen-Ion Concentration; D008051:Lipid Bilayers; D008081:Liposomes; D009682:Magnetic Resonance Spectroscopy; D008561:Membrane Fusion; D008563:Membrane Lipids / Q000737:chemistry*; D010712:Phosphatidic Acids / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry*
D008051:Lipid Bilayers; D008081:Liposomes; D008563:Membrane Lipids; D010712:Phosphatidic Acids; D010743:Phospholipids; D002118:Calcium