Deglycosylation of fibrinogen accelerates polymerization and increases lateral aggregation of fibrin fibers.
Fibrinogen, the major structural precursor of blood clots, was deglycosylated by peptide-N-(N-acetyl-beta-glucosaminyl)asparagine amidase without denaturation of the polypeptide chains. Deglycosylated fibrinogen behaved normally in clinical coagulation assays, although it is less soluble than normal fibrinogen. However, the turbidity of clots formed from deglycosylated fibrinogen always rose faster and higher than that of clots from normal fibrinogen. Scanning and transmission electron microscopy demonstrated that fibrin made from clots of deglycosylated fibrinogen consisted of thicker, less-branched fiber bundles in a more porous network. Moreover, the degree of lateral aggregation was directly related to clot turbidity and inversely related to branching. Deglycosylation promoted turbidity development, lateral aggregation, and porosity of clots under all conditions tested. All other steps in the coagulation pathways appeared to be unaffected by the absence of carbohydrate. These results suggest that carbohydrate constitutively affects the behavior of deglycosylated fibrinogens by 1) contributing a repulsive force that promotes fibrinogen solubility and limits fibrin assembly and 2) sensitizing fibrin to conditions that influence assembly and clot structure.
Langer
B G
BG
Department of Medicine, Johns Hopkins University, Baltimore, Maryland 21205.
Weisel
J W
JW
Dinauer
P A
PA
Nagaswami
C
C
Bell
W R
WR
D005174:Factor XII / Q000378:metabolism; D005177:Factor XIII Deficiency / Q000097:blood; D005337:Fibrin / Q000378:metabolism* / Q000648:ultrastructure; D005340:Fibrinogen / Q000302:isolation & purification / Q000378:metabolism* / Q000648:ultrastructure; D006031:Glycosylation; D006801:Humans; D007700:Kinetics; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D008958:Models, Molecular; D013917:Thrombin / Q000378:metabolism
D046911:Macromolecular Substances; D005174:Factor XII; D005337:Fibrin; D005340:Fibrinogen; D013917:Thrombin