Early cross-linked fibrin in human plasma contains alpha-polymers with intact fibrinopeptide A.
alpha-polymer formation, as opposed to gamma-chain dimerization has been considered a relatively late event in factor XIII-induced fibrin stabilization. Recently it has been shown, however, that plasma from healthy individuals and from patients with fibrinaemia contains small amounts of soluble fibrin/fibrinogen oligomers interlinked through dimerized gamma-chains as well as cross-linked alpha-chains. The present work was carried out to see if these early alpha-chain polymers also arise during coagulation of plasma in vitro. Plasma samples from healthy individuals, prepared by immediate centrifugation of blood collected without anticoagulant, were allowed to clot spontaneously for varying periods. The plasma clots were solubilized in SDS-urea-mercaptoethanol and samples were subjected to SDS-PAGE and Western blotting using polyclonal antibodies to human fibrinogen, or monoclonal antibodies specific either for A alpha/alpha-chains, for fibrinopeptide A-containing chains, for the N-terminus of the fibrin beta-chain or for the gamma-chains. Fibrin/fibrinogen oligomers were seen to form long before visible gelation of plasma. These oligomers were cross-linked through gamma-chain dimerization, but also through A alpha- or alpha-chain polymerization. The number and amount of alpha-polymers containing A alpha-chains increased immediately after clot formation, but these disappeared about 20 min later, due to complete removal of fibrinopeptide A (FPA) by thrombin. It is concluded that alpha-polymer formation is a very early event during plasma coagulation in vitro, and that both A alpha- and alpha-chains are involved.
Grøn
B
B
Research Institute for Internal Medicine, University of Oslo, Rikshospitalet, Norway.
Filion-Myklebust
C
C
Bennick
A
A
Nieuwenhuizen
W
W
Matsueda
G R
GR
Brosstad
F
F
D001777:Blood Coagulation; D001797:Blood Protein Electrophoresis; D015153:Blotting, Western; D005176:Factor XIII / Q000378:metabolism; D005337:Fibrin / Q000378:metabolism; D005344:Fibrinopeptide A / Q000032:analysis*; D006801:Humans; D008970:Molecular Weight; D011108:Polymers
D011108:Polymers; D005344:Fibrinopeptide A; D005337:Fibrin; D005176:Factor XIII