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(Received for publication, January 5, 1996, and in revised form, August 12, 1996)
From the Grupo Biomembranas (Unidad Asociada al Cosejo Superior de
Investigaciones Ciéntifícas), Departamento de
Bioquímica, Universidad del País Vasco, Apartado 644,
48080 Bilbao, Spain
When large unilamellar vesicles consisting of
sphingomyelin:phosphatidylethanolamine:cholesterol (2:1:1 molar ratio)
are treated with sphingomyelinase, production of ceramides in the
bilayer is accompanied by leakage of vesicle aqueous contents and by
vesicle aggregation in the absence of lipid mixing or vesicle fusion.
This is in contrast to the situation of
phosphatidylcholine:phosphatidylethanolamine:cholesterol (2:1:1
molar ratio) liposomes when treated with phospholipase C. In that case,
in situ generation of diacylglycerol leads to vesicle
aggregation followed by vesicle fusion in the absence of leakage
(Nieva, J. L., Goñi, F. M., and Alonso, A. (1989)
Biochemistry 28, 7364-7367). Moreover, when ceramides
(5-10 mol %) are included in the formulation of the
phosphatidylcholine-containing vesicles, they reduce the lag time of
phospholipase C-induced fusion, although they are less active than
diacylglycerols in this respect. 31P NMR studies of aqueous
lipid dispersions show that diacylglycerols as well as ceramides induce
a thermotropic lamellar to non-lamellar phase transition in both
phospholipid:cholesterol mixtures under study although
sphingomyelin-containing bilayers are more stable than those containing
phosphatidylcholine, and ceramide is less active than diacylglycerol in
promoting non-lamellar phase formation. These observations are relevant
to both the physiological role of ceramides and the current views on
the mechanism of membrane fusion.
Volume 271, Number 43,
Issue of October 25, 1996
pp. 26616-26621
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
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