Interaction between phosphatidylserine and the isolated cytoskeleton of human blood platelets.
Binding experiments were performed to demonstrate a direct interaction between cytoskeletons from human blood platelets and phosphatidylserine. A centrifugation technique using radiolabeled phosphatidylserine-vesicles and Triton X-100 insoluble residues from unstimulated human platelets was used to assess the binding. Interaction between cytoskeleton and phospholipid is demonstrated to be specific for phosphatidylserine. No binding was observed for phosphatidylcholine. The binding of phosphatidylserine was saturable and dependent on the concentration of cytoskeleton used. The interaction between phosphatidylserine and the cytoskeleton appeared to be completely reversible. The existence of a reversible and specific interaction between phosphatidylserine and the cytoskeleton of unstimulated platelets would suggest a role for the cytoskeleton in the maintenance of the asymmetric distribution of this lipid in the plasma membrane. We have previously shown (Comfurius et al. (1985) Biochim. Biophys. Acta 815, 143-148) that in activated platelets a strong correlation exists between degradation of platelet cytoskeletal proteins by the endogenous calcium-dependent proteinase (calpain) and exposure of phosphatidylserine at their outer surface. Nevertheless, hydrolysis of the isolated cytoskeleton by calpain did not result in a change in the parameters of the binding between phosphatidylserine and cytoskeleton. Also, sulfhydryl oxidation of the cytoskeleton by diamide did not affect its binding properties for phosphatidylserine, in spite of the fact that diamide treatment of platelets results in exposure of phosphatidylserine at the outer surface. Exposition of phosphatidylserine upon activation of platelets cannot be directly ascribed to a change in affinity or number of binding sites of the modified cytoskeleton as measured in model systems. However, it cannot be excluded that topological rearrangements of the cytoskeleton as occur within the cell during platelet activation lead to a decreased contact between cytoskeleton and lipid, irrespective of the binding parameters.
Comfurius
P
P
Department of Biochemistry, University of Limburg, Maastricht, The Netherlands.
Bevers
E M
EM
Zwaal
R F
RF
D001665:Binding Sites; D001792:Blood Platelets / Q000378:metabolism*; D002154:Calpain / Q000494:pharmacology; D002458:Cell Fractionation; D003599:Cytoskeleton / Q000378:metabolism*; D003958:Diamide / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008563:Membrane Lipids / Q000378:metabolism; D010718:Phosphatidylserines / Q000378:metabolism*
D008563:Membrane Lipids; D010718:Phosphatidylserines; D003958:Diamide; D002154:Calpain