Transglutaminase-catalyzed crosslinking of the Aα and γ constituent chains in fibrinogen

Studies on transglutaminases usually focus on the polymerization of protein substrates by intermolecular N-epsilon(gamma-glutamyl)lysine bridges, without considering the possibility that the monomeric protein units, themselves, could also become crosslinked internally. Both types of crosslinks are produced in the reaction of fibrinogen with red cell transglutaminase, We isolated the transglutaminase-modif led, mostly monomeric form (92-96 %) of fibrinogen with a N-epsilon(gamma-glutamyl)lysine content of approximate to 1.6 moles/mole of fibrinogen. The preparation was fully clottable by thrombin, but the rates of release of fibrinopeptides and clotting times were delayed compared with control. Hybrid A alpha.gamma type of crosslinking, the hallmark of the reaction of the transglutaminase with fibrinogen, occurred by bridging the A alpha(408-421) chain segment of the protein to that of gamma(392-406). Rotary shadowed electron microscope images showed many monomers to be bent, and the crosslinks seemed to hind the otherwise flexible alpha C domain closer to the backbone of fibrinogen.