title: Loss of membrane phospholipid asymmetry in platelets and red cells may be associated with calcium-induced shedding of plasma membrane and inhibition of aminophospholipid translocase.
abstract: Influx of calcium in platelets and red cells produces formation of vesicles shed from the plasma membrane. The time course of the shedding process closely correlates with the ability of both cells to stimulate prothrombinase activity when used as a source of phospholipid in the prothrombinase assay. This reflects increased surface exposure of phosphatidylserine, presumably resulting from a loss in membrane asymmetry. Evidence is presented that the shed vesicles have a random phospholipid distribution, while the remnant cells show a progressive loss of membrane phospholipid asymmetry when more shedding occurs. Removal of intracellular calcium produces a decrease of procoagulant activity of the remnant cells but not of that of the shed vesicles. This is consistent with reactivation of aminophospholipid translocase activity, being first inhibited by intracellular calcium and subsequently reactivated upon calcium removal. Involvement of aminophospholipid translocase is further supported by the observation that reversibility of procoagulant activity is also dependent on metabolic ATP and reduced sulfhydryl groups. The finding that this reversibility process is not apparent in shed vesicles may be ascribed to the absence of translocase or to a lack of ATP. These data support and extend the suggestion made by Sims et al. [1989) J. Biol. Chem. 264, 17049-17057) that membrane fusion, which is required for shedding to occur, produces transient flip-flop sites for membrane phospholipids. Furthermore, the present results indicate that scrambling of membrane phospholipids can only occur provided that aminophospholipid translocase is inactive.
authors.affiliation: Department of Biochemistry, University of Limburg, Maastricht, The Netherlands.
authors.forename: P
authors.initials: P
authors.lastname: Comfurius
authors.forename: J M
authors.initials: JM
authors.lastname: Senden
authors.forename: R H
authors.initials: RH
authors.lastname: Tilly
authors.forename: A J
authors.initials: AJ
authors.lastname: Schroit
authors.forename: E M
authors.initials: EM
authors.lastname: Bevers
authors.forename: R F
authors.initials: RF
authors.lastname: Zwaal
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D001792:Blood Platelets / Q000502:physiology* / Q000648:ultrastructure; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002154:Calpain / Q000037:antagonists & inhibitors; D002352:Carrier Proteins / Q000378:metabolism*; D002451:Cell Compartmentation; D004533:Egtazic Acid / Q000494:pharmacology; D004789:Enzyme Activation; D004910:Erythrocyte Membrane / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D066298:In Vitro Techniques; D008563:Membrane Lipids / Q000502:physiology*; D008565:Membrane Proteins / Q000378:metabolism*; D010718:Phosphatidylserines / Q000378:metabolism; D049294:Phospholipid Transfer Proteins; D010743:Phospholipids / Q000502:physiology*; D015539:Platelet Activation; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013925:Thromboplastin / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D008563:Membrane Lipids; D008565:Membrane Proteins; D010718:Phosphatidylserines; D049294:Phospholipid Transfer Proteins; D010743:Phospholipids; D013438:Sulfhydryl Compounds; D000001:Calcimycin; D004533:Egtazic Acid; D000255:Adenosine Triphosphate; D013925:Thromboplastin; D002154:Calpain; D002118:Calcium