Phospholipase A2 promotes raft budding and fission from giant liposomes.

Cellular processes involving membrane vesiculation are related to cellular transport and membrane components trafficking. Endocytosis, formation of caveolae and caveosomes, as well as Golgi membranes traffic have been linked to the existence and dynamics of particular types of lipid/protein membrane domains, enriched in sphingolipids and cholesterol, called rafts [Nature 387 (1997) 569; Trends Cell Biol. 12 (2002) 296; Biochemistry 27 (1988) 6197]. In addition, the participation of phospholipases in the vesiculation of Golgi and other membranes has been already established [Traffic 1 (2000) 504] essentially in their role in the production of second messenger molecules. In this work we illustrate with raft-containing giant lipid vesicles a mechanism for raft-vesicle expulsion from the membrane due to the activity of a single enzyme-phospholipase A(2) (PLA(2)). This leads to the hypothesis that the PLA(2), apart from its role in second messenger generation, might play a direct and general role in the vesiculation processes underlying the intermembrane transport of rafts through purely physicochemical mechanisms. These mechanisms would be: enzyme adsorption leading to membrane curvature generation (budding), and enzyme activity modulation of the line tension at the raft boundaries, which induces vesicle fission.

[1]  P. Luisi,et al.  Microinjection into giant vesicles and light microscopy investigation of enzyme-mediated vesicle transformations. , 1996, Chemistry & biology.

[2]  G. Karlström,et al.  Phase equilibria in the phosphatidylcholine-cholesterol system. , 1987, Biochimica et biophysica acta.

[3]  M. Angelova,et al.  Interactions of DNA with giant liposomes. , 1999, Chemistry and physics of lipids.

[4]  P. de Figueiredo,et al.  Phospholipase A2 Antagonists Inhibit Constitutive Retrograde Membrane Traffic to the Endoplasmic Reticulum , 2000, Traffic.

[5]  B. de Kruijff,et al.  Effects of lysophosphatidylcholines on phosphatidylcholine and phosphatidylcholine/cholesterol liposome systems as revealed by 31P-NMR, electron microscopy and permeability studies. , 1981, Biochimica et biophysica acta.

[6]  O. G. Mouritsen,et al.  Biophysical mechanisms of phospholipase A2 activation and their use in liposome‐based drug delivery , 2002, FEBS letters.

[7]  P. Quinn,et al.  Ceramides increase the activity of the secretory phospholipase A2 and alter its fatty acid specificity. , 2002, The Biochemical journal.

[8]  C. Casals,et al.  Differential partitioning of pulmonary surfactant protein SP-A into regions of monolayers of dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylcholine/dipalmitoylphosphatidylglycerol. , 1998, Biophysical journal.

[9]  B. de Kruijff,et al.  Visualizing detergent resistant domains in model membranes with atomic force microscopy , 2001, FEBS letters.

[10]  R. Pagano Lipid traffic in eukaryotic cells: mechanisms for intracellular transport and organelle-specific enrichment of lipids. , 1990, Current opinion in cell biology.

[11]  M. Angelova,et al.  Giant Vesicles: Imitating the Cytological Processes of Cell Membranes , 1998 .

[12]  P. Kinnunen,et al.  Vectorial budding of vesicles by asymmetrical enzymatic formation of ceramide in giant liposomes. , 2000, Biophysical journal.

[13]  D. Zhelev,et al.  Lysolipid exchange with lipid vesicle membranes , 1995, Annals of Biomedical Engineering.

[14]  P. Kinnunen,et al.  Macroscopic consequences of the action of phospholipase C on giant unilamellar liposomes. , 2002, Biophysical journal.

[15]  E. Ikonen,et al.  Functional rafts in cell membranes , 1997, Nature.

[16]  A. Dautry‐Varsat,et al.  Enhancement of endocytosis due to aminophospholipid transport across the plasma membrane of living cells. , 1999, The American journal of physiology.

[17]  E Gratton,et al.  Lipid rafts reconstituted in model membranes. , 2001, Biophysical journal.

[18]  R. Lipowsky Domains and Rafts in Membranes – Hidden Dimensions of Selforganization , 2002, Journal of biological physics.

[19]  F. G. van der Goot,et al.  Oiling the wheels of the endocytic pathway. , 2002, Trends in cell biology.

[20]  E. London,et al.  Interactions between saturated acyl chains confer detergent resistance on lipids and glycosylphosphatidylinositol (GPI)-anchored proteins: GPI-anchored proteins in liposomes and cells show similar behavior. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[21]  P. Quinn,et al.  Cholesterol relieves the inhibitory effect of sphingomyelin on type II secretory phospholipase A2. , 1998, The Biochemical journal.

[22]  Petra Schwille,et al.  Probing Lipid Mobility of Raft-exhibiting Model Membranes by Fluorescence Correlation Spectroscopy* , 2003, Journal of Biological Chemistry.

[23]  R. Bittman,et al.  Interaction of cholesterol with sphingomyelin in monolayers and vesicles. , 1994, Biochemistry.

[24]  O. Mouritsen Theoretical models of phospholipid phase transitions. , 1991, Chemistry and physics of lipids.

[25]  E. Ikonen,et al.  Roles of lipid rafts in membrane transport. , 2001, Current opinion in cell biology.

[26]  C. W. M. ADAMS,et al.  Chemistry and Physics of Lipids , 1967, Nature.

[27]  M. Martín,et al.  Protein kinase A activity is required for the budding of constitutive transport vesicles from the trans-Golgi network. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[28]  C. Casals,et al.  Pulmonary surfactant protein A interacts with gel-like regions in monolayers of pulmonary surfactant lipid extract. , 2000, Biophysical journal.

[29]  D. S. Dimitrov,et al.  A mechanism of liposome electroformation , 1988 .

[30]  W. Lehmann,et al.  Evidence for Segregation of Sphingomyelin and Cholesterol during Formation of Copi-Coated Vesicles , 2000, The Journal of cell biology.

[31]  D. Sabatini,et al.  The production of post-Golgi vesicles requires a protein kinase C-like molecule, but not its phosphorylating activity , 1996, The Journal of cell biology.

[32]  R. Parton,et al.  Sphingolipid transport from the trans‐Golgi network to the apical surface in permeabilized MDCK cells , 1992, FEBS letters.

[33]  G van Meer,et al.  Lipid sorting in epithelial cells. , 1988, Biochemistry.

[34]  H. Gaub,et al.  Atomic force microscope imaging of phospholipid bilayer degradation by phospholipase A2. , 1998, Biophysical journal.