Identification of covalently linked trimeric and tetrameric D domains in crosslinked fibrin.
Following proteolytic conversion of fibrinogen to fibrin, clot assembly commences with formation of double-stranded fibrils that subsequently branch extensively in forming a three-dimensional network. Plasmin digests of fibrin clots that had first been covalently crosslinked by plasma transglutaminase (factor XIIIa) contained multimeric proteolytic fragments composed of crosslinked outer (D) domains of neighboring fibrin molecules. Two of these were larger than the well-known "D dimer" fragment and corresponded to D trimers and D tetramers, respectively. Whereas D dimers originate from crosslinked D domains at bimolecular junctions within two-stranded fibrils, D trimers and D tetramers evidently arise through crosslinking of contiguous D domains at trimolecular and tetramolecular junctions or at fibril branch points, respectively. Measurement of the widths of fibrils comprising trifunctional branches in thin fiber networks revealed tetramolecular branch points, which are formed by bifurcation of two double-stranded fibrils. In addition, another type of trifunctional structure, which we term the trimolecular branch point, was composed of three double-stranded fibrils. Crosslinking of D domains to form trimers may occur at this type of junction. These findings add to our understanding of the crosslinking arrangements that stabilize fibrin clot structure and the ways that fibrin molecules polymerize to form branches in the clot matrix.
Mosesson
M W
MW
University of Wisconsin Medical School, Sinai Samaritan Medical Center, Milwaukee 53233.
Siebenlist
K R
KR
Amrani
D L
DL
DiOrio
J P
JP
D003432:Cross-Linking Reagents; D005337:Fibrin / Q000378:metabolism* / Q000648:ultrastructure; D005340:Fibrinogen / Q000378:metabolism / Q000648:ultrastructure; D005341:Fibrinolysin / Q000378:metabolism; D006801:Humans; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008962:Models, Theoretical
D003432:Cross-Linking Reagents; D046911:Macromolecular Substances; D005337:Fibrin; D005340:Fibrinogen; D005341:Fibrinolysin