ATP-dependent fusion of liposomes with the Golgi apparatus of perforated cells.
We examined the interactions of lipid vesicles (liposomes) labeled with various fluorescent markers with the intracellular membranes of semi-intact ("perforated") fibroblasts. When incubations were performed in the presence of an ATP-regenerating system, both vesicle lipids and entrapped water soluble markers were transferred to the Golgi apparatus of treated cells, indicative of membrane fusion. Fusion occurred using unilamellar vesicles 30-80 nm in diameter and composed of phosphatidylcholine alone, but was inhibited when equimolar amounts of either phosphatidylserine or sphingomyelin were present in the vesicles. Lipid vesicle-Golgi membrane fusion was also inhibited by pretreatment of the perforated cells with N-ethylmaleimide. These findings suggest that lipid vesicles may be useful for delivery of labeled lipids, macromolecules, and dyes to the Golgi apparatus, and for modeling the interactions of transport vesicles with the Golgi apparatus.
Kobayashi
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Department of Embryology, Carnegie Institution of Washington, Baltimore, Maryland 21210.
Pagano
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D000255:Adenosine Triphosphate / Q000502:physiology*; D001692:Biological Transport; D002451:Cell Compartmentation; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D002518:Ceramides; D006056:Golgi Apparatus / Q000502:physiology*; D006801:Humans; D007425:Intracellular Membranes / Q000502:physiology*; D008081:Liposomes; D008561:Membrane Fusion; D008563:Membrane Lipids / Q000502:physiology; D008856:Microscopy, Fluorescence; D013329:Structure-Activity Relationship
D002518:Ceramides; D008081:Liposomes; D008563:Membrane Lipids; D000255:Adenosine Triphosphate