Plasminogen associates with phosphatidylserine-exposing platelets and contributes to thrombus lysis under flow.

Abstract

The interaction of plasminogen with platelets and their localization during thrombus formation and fibrinolysis under flow are not defined. Using a novel model of whole blood thrombi, formed under flow, we examine dose-dependent fibrinolysis using fluorescence microscopy. Fibrinolysis was dependent upon flow and the balance between fibrin formation and plasminogen activation, with tissue plasminogen activator-mediated lysis being more efficient than urokinase plasminogen activator-mediated lysis. Fluorescently labeled plasminogen radiates from platelet aggregates at the base of thrombi, primarily in association with fibrin. Hirudin attenuates, but does not abolish plasminogen binding, denoting the importance of fibrin. Flow cytometry revealed that stimulation of platelets with thrombin/convulxin significantly increased the plasminogen signal associated with phosphatidylserine (PS)-exposing platelets. Binding was attenuated by tirofiban and Gly-Pro-Arg-Pro amide, confirming a role for fibrin in amplifying plasminogen binding to PS-exposing platelets. Confocal microscopy revealed direct binding of plasminogen and fibrinogen to different platelet subpopulations. Binding of plasminogen and fibrinogen co-localized with PAC-1 in the center of spread platelets. In contrast, PS-exposing platelets were PAC-1 negative, and bound plasminogen and fibrinogen in a protruding "cap." These data show that different subpopulations of platelets harbor plasminogen by diverse mechanisms and provide an essential scaffold for the accumulation of fibrinolytic proteins that mediate fibrinolysis under flow.

• Authors: Claire S Whyte, Frauke Swieringa, Tom G Mastenbroek, Ausra S Lionikiene, Marcus D Lancé, Paola E J van der Meijden, Johan W M Heemskerk, Nicola J Mutch
• Journal: Blood (Blood) Vol. 125 Issue 16 Pg. 2568-78 (Apr 16 2015) ISSN: 1528-0020 [Electronic] United States
• PMID: 25712989 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2015 by The American Society of Hematology.

Chemical References

• Crotalid Venoms
• Fibrinolytic Agents
• Hemostatics
• Hirudins
• Lectins, C-Type
• Oligopeptides
• Phosphatidylserines
• convulxin
• Tyrosine
• glycyl-prolyl-arginyl-proline
• Fibrin
• Thrombin
• Tissue Plasminogen Activator
• Fibrinolysin
• Urokinase-Type Plasminogen Activator
• Tirofiban

Topics

• Blood Platelets (drug effects, metabolism)
• Crotalid Venoms (pharmacology)
• Fibrin (metabolism)
• Fibrinolysin (metabolism)
• Fibrinolysis (drug effects)
• Fibrinolytic Agents (pharmacology)
• Flow Cytometry
• Hemostatics (pharmacology)
• Hirudins (pharmacology)
• Lectins, C-Type
• Microscopy, Confocal
• Oligopeptides (pharmacology)
• Phosphatidylserines (metabolism)
• Protein Binding (drug effects)
• Rheology
• Shear Strength
• Thrombin (pharmacology)
• Thrombosis (metabolism)
• Tirofiban
• Tissue Plasminogen Activator (metabolism)
• Tyrosine (analogs & derivatives, pharmacology)
• Urokinase-Type Plasminogen Activator (metabolism)