Factor XIIIa binding to activated platelets is mediated through activation of glycoprotein IIb-IIIa.
Stabilization of a clot is dependent on fibrin cross-linking mediated by the transglutaminase, factor XIIIa (FXIIIa). In addition to fibrin stabilization, FXIIIa acts on a number of platelet-reactive proteins, including fibronectin and vitronectin, as well as the platelet proteins, glycoprotein (GP) IIb-IIIa, myosin, and actin. However, conditions inducing the platelet-activation dependent binding of FXIIIa have not been characterized nor have the sites mediating FXIIIa binding been identified. The generation of FXIIIa and consequent detection of FXIIIa on the platelet surface were compared with other thrombin-induced activation events; the rate at which FXIIIa bound to activated platelets was much slower than platelet degranulation or fibrin(ogen) binding. Whereas platelets could be rapidly induced to express a functional receptor for FXIIIa, the rate of FXIIIa binding to platelets is limited by the rate of conversion of FXIII to FXIIIa. Immunoprecipitation of radiolabeled platelets using polyclonal anti-FXIII A-chain antibody identified two proteins corresponding to GPIIb and GPIIIa. Preincubation of intact platelets with 7E3, a monoclonal antibody that blocks the fibrinogen binding site, or GRGDSP peptide inhibited FXIIIa binding by about 95% when measured by flow cytometry; FXIIIa binding to purified GPIIb-IIIa was also inhibited by 7E3. The binding of FXIIIa to purified GPIIb-IIIa was enhanced by the addition of fibrinogen, but not by that of fibronectin or thrombospondin, suggesting that FXIIIa also binds to fibrinogen associated with the complex. These observations suggest that activated platelets bearing FXIIIa may enhance stabilization of platelet-rich thrombi through surface-localized cross-linking events.
Cox
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Department of Pathology, University of British Columbia, Vancouver, Canada.
Devine
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D000244:Adenosine Diphosphate / Q000494:pharmacology; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D015703:Antigens, CD / Q000032:analysis; D016718:Arachidonic Acid / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D000001:Calcimycin / Q000494:pharmacology; D003094:Collagen / Q000494:pharmacology; D004837:Epinephrine / Q000494:pharmacology; D005340:Fibrinogen / Q000494:pharmacology; D005353:Fibronectins / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008562:Membrane Glycoproteins / Q000494:pharmacology; D008565:Membrane Proteins / Q000097:blood / Q000302:isolation & purification; D009842:Oligopeptides / Q000494:pharmacology*; D015539:Platelet Activation; D010980:Platelet Membrane Glycoproteins / Q000302:isolation & purification / Q000378:metabolism*; D012016:Reference Values; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013915:Thrombasthenia / Q000097:blood*; D013917:Thrombin / Q000494:pharmacology; D019699:Thrombospondins; D013997:Time Factors; D011503:Transglutaminases / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D005353:Fibronectins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009842:Oligopeptides; D010980:Platelet Membrane Glycoproteins; D019699:Thrombospondins; D016718:Arachidonic Acid; D000001:Calcimycin; D000244:Adenosine Diphosphate; C047981:arginyl-glycyl-aspartic acid; D005340:Fibrinogen; D003094:Collagen; C058161:glycyl-arginyl-glycyl-aspartyl-seryl-proline; D011503:Transglutaminases; D013917:Thrombin; D013755:Tetradecanoylphorbol Acetate; D004837:Epinephrine