Granulocyte-platelet interactions and platelet fibrinogen receptor exposure.
We have examined the interaction of human granulocyte elastase with human platelets. Incubation of human platelets with human granulocyte elastase exposed active fibrinogen-binding sites as evidenced by 125I-labeled fibrinogen binding and spontaneous fibrinogen-induced platelet aggregation. The aggregation of platelets by fibrinogen occurred at low concentrations of human granulocyte elastase (0.5-1 microgram/ml). Platelets pretreated with human granulocyte elastase exposed an average of 10,500 fibrinogen binding sites per platelet, i.e., about one-third the number of binding sites exposed by optimal concentrations of ADP. With the use of a polyclonal antiplatelet membrane antibody, the glycoproteins IIb (GPIIb), IIIa (GPIIIa), and a 60,000-Da (60 kDa) protein (66 kDa in a reduced system) derived from GPIIIa were immunoprecipitated from the surface of detergent extracts of human 125I-radiolabeled platelets pretreated with increasing concentrations of human granulocyte elastase. Experiments performed by immunoblotting with use of polyclonal and monoclonal antibodies directed to GPIIIa showed that pretreatment of human platelets with granulocyte elastase resulted in the appearance of an additional proteolytic derivative of GPIIIa migrating with an apparent molecular mass of 120 kDa in a nonreduced system. GPIIIa appears to be the preferred substrate of elastase, since GPIIb was not degraded by human granulocyte elastase. We conclude that 1) the proteolytic action of human granulocyte elastase on platelet GPIIIa results in the formation of two major hydrolytic products, and 2) human granulocyte elastase exposes active fibrinogen-binding sites associated with the GPIIb/GPIIIa complex, resulting in direct platelet aggregation by fibrinogen.
Department of Psychiatry, University of Vermont, Burlington 05405.
E
E
Kornecki
Y H
YH
Ehrlich
R
R
Egbring
M
M
Gramse
R
R
Seitz
A
A
Eckardt
H
H
Lukasiewicz
S
S
Niewiarowski
D000255:Adenosine Triphosphate / Q000097:blood; D001792:Blood Platelets / Q000502:physiology*; D005340:Fibrinogen / Q000502:physiology*; D006098:Granulocytes / Q000502:physiology*; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight; D010196:Pancreatic Elastase / Q000097:blood / Q000302:isolation & purification; D010974:Platelet Aggregation; D010976:Platelet Count; D010980:Platelet Membrane Glycoproteins / Q000302:isolation & purification / Q000502:physiology*
D010980:Platelet Membrane Glycoproteins; D000255:Adenosine Triphosphate; D005340:Fibrinogen; D010196:Pancreatic Elastase