The (DD)E complex is maintained by a composite fibrin polymerization site.
The (DD)E complex is the major cross-linked fibrin degradation fragment. Structural components required for maintenance of the (DD)E complex were examined in order to better understand clot structure and the contribution of specific polypeptide chain segments in the process of polymerization. First, the (DD)E complex was reversibly dissociated by peptides derived from the alpha-chain NH2-terminus of fibrin having a minimal sequence of GPR (alpha 17-19). In addition, the complex was partially dissociated by peptide beta 40-54, while beta 50-55 and peptides derived from the fibrin beta-chain NH2-terminus had no effect. Second, monoclonal antibody (mAb) 1B6, specific for the alpha-chain NH2-terminus of fibrin, reacted rapidly with fragment E1, but did not recognize the corresponding epitope on the (DD)E complex. On the other hand, mAb 59D8, specific for GHRPL at the beta-chain NH2-terminus of fibrin, reacted with the (DD)E complex in a dose-dependent manner. Third, the (DD)E complex was irreversibly dissociated by proteolytic cleavage of fragment E1 by either thrombin, which removed GPR from the alpha-chain NH2-terminus, or Crotalus atrox protease III, which released beta 15-42. It has been concluded that fragment E1 contains a composite polymerization site consisting at least of residues alpha 17-19 and beta 20-49, which together maintain the (DD)E complex. These results illustrate that the complex is kept together by complementary binding sites which form a nucleus of linear fibrin polymerization sites. The (DD)E complex can thus be considered as a soluble model of fibrin clot.(ABSTRACT TRUNCATED AT 250 WORDS)
Moskowitz
K A
KA
Department of Biochemistry, Temple University School of Medicine, Philadelphia, Pennsylvania 19140.
Budzynski
A Z
AZ
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D001667:Binding, Competitive; D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents; D003435:Crotalid Venoms / Q000201:enzymology; D017839:Crotalus; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005337:Fibrin / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D005455:Fluorescent Antibody Technique; D008666:Metalloendopeptidases / Q000737:chemistry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D011108:Polymers; D013917:Thrombin / Q000737:chemistry
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D003435:Crotalid Venoms; D000939:Epitopes; D010446:Peptide Fragments; D011108:Polymers; D005337:Fibrin; D013917:Thrombin; D008666:Metalloendopeptidases; C046583:pitrilysin; D016650:Fluorescein-5-isothiocyanate