title: gammaA/gamma' fibrinogen inhibits thrombin-induced platelet aggregation.
abstract: The minor gammaA/gamma' fibrinogen isoform contains a high affinity binding site for thrombin exosite II that is lacking in the major gammaA/gammaA fibrinogen isoform. We therefore investigated the biological consequences of the gamma' chain binding to thrombin. Thrombin-induced platelet aggregation was inhibited by gammaA/gamma' fibrinogen. Carboxyl terminal peptide fragment gamma'410-427 from the gamma' chain was also inhibitory, with an IC(50) of approximately 200 microM in whole plasma. Deletion of the peptide from either the amino or carboxyl end significantly decreased inhibition. In contrast to thrombin-induced platelet aggregation, aggregation induced by epinephrine, ADP, arachidonic acid, or SFLLRN peptide showed little inhibition by the gamma' peptide. The inhibition of thrombin-induced platelet aggregation was not due to direct inhibition of the thrombin active site, since cleavage of a small peptidyl substrate was 91% of normal even in the presence of 1 mM gamma'410-427. The gamma'410-427 peptide blocked platelet adhesion to immobilized thrombin under both static and flow conditions, blocked soluble thrombin binding to platelet GPIbalpha, and inhibited PAR1 cleavage by thrombin. These results suggest that the gamma' chain of fibrinogen inhibits thrombin-induced platelet aggregation by binding to thrombin exosite II. Thrombin that is bound to the gamma' chain is thereby prevented from activating platelets, while retaining its amidolytic activity.
authors.affiliation: Department of Biomedical Sciences, Missouri State University, Springfield, MO, USA.
authors.forename: Rehana S
authors.initials: RS
authors.lastname: Lovely
authors.forename: Chantelle M
authors.initials: CM
authors.lastname: Rein
authors.forename: Tara C
authors.initials: TC
authors.lastname: White
authors.forename: Sari A
authors.initials: SA
authors.lastname: Jouihan
authors.forename: Lynn K
authors.initials: LK
authors.lastname: Boshkov
authors.forename: Antony C
authors.initials: AC
authors.lastname: Bakke
authors.forename: Owen J
authors.initials: OJ
authors.lastname: McCarty
authors.forename: David H
authors.initials: DH
authors.lastname: Farrell
mesh_terms: D001792:Blood Platelets / Q000378:metabolism*; D005340:Fibrinogen / Q000378:metabolism*; D006801:Humans; D010446:Peptide Fragments / Q000378:metabolism*; D010973:Platelet Adhesiveness; D010974:Platelet Aggregation; D010979:Platelet Function Tests; D011485:Protein Binding; D044463:Receptor, PAR-1 / Q000378:metabolism; D013917:Thrombin / Q000378:metabolism*; D013997:Time Factors
chemical_list: D010446:Peptide Fragments; D044463:Receptor, PAR-1; C027539:fibrinopeptides gamma; D005340:Fibrinogen; D013917:Thrombin