Vesiculation of platelet plasma membranes. Dilauroylglycerophosphocholine-induced shedding of a platelet plasma membrane fraction enriched in acetylcholinesterase activity.
Incubation of washed rabbit platelets with suspensions of dilauroylglycerophosphocholine resulted in the shedding of vesicles without causing any appreciable leakage of cytoplasmic marker (lactate dehydrogenase) or organelle marker [( 14C]serotonin). The response was dependent on incubation time, concentration of dilauroylglycerophosphocholine and reaction temperature. Vesicles were separated from platelets and exogenous dilauroylglycerophosphocholine by a series of centrifugation steps. An average diameter of vesicles was 100-200 nm on scanning electron microscopy. Vesicles were enriched 5-fold in plasma membrane marker enzyme, acetylcholinesterase, whereas specific activities of lactate dehydrogenase and intracellular membrane marker enzyme, NADH-cytochrome c reductase were decreased in vesicles. Protein analysis by SDS-polyacrylamide gel electrophoresis showed that actin and actin-binding protein were present, while myosin was barely detectable in vesicles. Vesicles contained all phospholipid species of intact platelets and cholesterol but almost 50% of phospholipids in vesicles was dilauroylglycerophosphocholine. The phospholipid to protein ratio in vesicles was about 6.5-times higher than in intact platelets.
D000110:Acetylcholinesterase / Q000378:metabolism*; D000818:Animals; D001792:Blood Platelets / Q000187:drug effects / Q000648:ultrastructure*; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000201:enzymology / Q000648:ultrastructure*; D002498:Centrifugation; D002784:Cholesterol / Q000097:blood; D008563:Membrane Lipids / Q000097:blood; D008565:Membrane Proteins / Q000097:blood; D008855:Microscopy, Electron, Scanning; D010713:Phosphatidylcholines / Q000494:pharmacology*; D010743:Phospholipids / Q000097:blood; D011817:Rabbits
D008563:Membrane Lipids; D008565:Membrane Proteins; D010713:Phosphatidylcholines; D010743:Phospholipids; C015153:1,2-dilauroylphosphatidylcholine; D002784:Cholesterol; D000110:Acetylcholinesterase