Summary
We previously described synthetic peptides of 19-21 amino acid residues, homologous
to the C-termini of fibrinogen Fib340 and Fib420, from the β-chain (Cβ), the extended αE chain (CαE) and near the end of the γ-chain
(preCγ) which elicited attachment (haptotactic) responses from mesenchymal cells.
We named these haptotactic peptides -Haptides. The effects of Haptides on fibrin clot
formation was evaluated and their possible effects on platelet aggregation was examined.
The Haptides Cβ, CαE and preCγ, (2-10 μM) increased fibrin clot turbidity and also
decreased thrombin-induced clotting time. Higher concentrations (>120 μM of Cβ or
preCγ) induced fibrinogen precipitation even without thrombin. These precipitates
exhibited different ultrastructure from thrombin-induced fibrin by scanning and transmission
microscopy. C-terminal peptides of the other fibrinogen chains exerted no such effects.
Sepharose beads covalently coated with either whole fibrinogen or Haptides (SB-Fib
or SB-Haptide) highly adsorbed free FITCHaptides. In aqueous solution, Haptides formed
nano-particles with average size of ∼150nm in diameter. We suggest that such positively
charged aggregates could serve to nucleate and accelerate fibrin gel formation. These
results also indicate that Cβ and preCγ sequences within fibrin(ogen) participate
in the docking and condensation of fibrin(ogen) during its assembly into a fibrin
clot. By contrast, Haptides up to 100µM did not bind to platelets, and had no effect
on platelet aggregation. Our findings highlight the roles of the C-terminal sequences
of the β and γ chains in fibrin(ogen) polymerization as well as in cell attachment.
Keywords
Fibrinogen - turbidity - Haptides - coagulation - aggregate