Factor XIII binds to the Aα- and Bβ- chains in the D-domain of fibrinogen: An immunoblotting study

Abstract

The binding sites in fibrinogen for Factor XIII were localized using an immunoblotting technique. Platelet Factor XIII bound to fibrinogen and to plasmin degradation products of fibrin(ogen) including Fragments: X, D 1–D 3, and D-dimer, but did not bind to Fragment E. Binding of Platelet Factor XIII was independent of calcium ions but could be inhibited by the presence of 0.5 M NaCl. Binding could also be inhibited by preincubating Factor XIII with a 100-fold molar excess of fibrinogen but not by 100-fold molar excess of Fragment E. Binding of Factor XIII to fibrinogen was specific, since several other proteins tested (ovalbumin, bovine serum albumin, α 2-macroglobulin, β-galactosidase, fructose kinase, lactic dehydrogenase, triose phosphate isomerase, fumarase and pyruvate kinase) did not bind Factor XIII. Furthermore, binding was not observed either when Factor XIII was left out or when antiFactor XIII antiserum was substituted with nonimmune serum. When fibrinogen was reduced prior to electrophoresis, Factor XIII bound to the Aα and Bβ chains of fibrinogen and des A,B fibrinogen, the Bβ-chain of Fragment X, but not the γ-chains. Localization of the Factor XIII binding sites to the carboxy terminal segments of the Aα and Bβ chains in the Fragment D-domain of fibrinogen could have important physiological consequences.