title: Isolation of a fibrin-binding fragment from blood coagulation factor XIII capable of cross-linking fibrin(ogen).
abstract: Purified platelet Factor XIII was radioiodinated and then partially degraded by thrombin or trypsin, and a fibrin-binding fragment was identified by autoradiography and immunoblotting following separation by SDS/polyacrylamide-gel electrophoresis. Limited proteolysis of 125I-Factor XIII by thrombin or trypsin produced an 125I-51 kDa fragment and an unlabelled 19 kDa fragment. The 51 kDa fragment was purified by h.p.l.c. on a TSK-125 gel-filtration column. Partial amino acid sequence analysis of the 51 kDa fragment indicated that it was similar in sequence to the Gly38-Lys513 segment in placental Factor XIII a-chain. More than 70% of the 51 kDa fragment bound to fibrin, whereas the 19 kDa fragment did not bind. The active site was localized to the 51 kDa fragment since this fragment expressed transglutaminase activity, cross-linked fibrin and fibrinogen and incorporated iodo[14C]acetamide into the active-site cysteine residue. Isolation of a fibrin-binding fragment expressing transglutaminase activity demonstrates that each a-chain of the dimeric Factor XIIIa could function independently to cross-link fibrin. The fibrin-binding site could play an important role in localizing Factor XIIIa to the fibrin clot.
authors.affiliation: Department of Medicine, Duke University Medical Center, Durham, NC 27710.
authors.forename: C S
authors.initials: CS
authors.lastname: Greenberg
authors.forename: J J
authors.initials: JJ
authors.lastname: Enghild
authors.forename: A
authors.initials: A
authors.lastname: Mary
authors.forename: J V
authors.initials: JV
authors.lastname: Dobson
authors.forename: K E
authors.initials: KE
authors.lastname: Achyuthan
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D003432:Cross-Linking Reagents; D005176:Factor XIII / Q000302:isolation & purification*; D005337:Fibrin / Q000378:metabolism*; D005340:Fibrinogen / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000302:isolation & purification*; D010447:Peptide Hydrolases
chemical_list: D003432:Cross-Linking Reagents; D010446:Peptide Fragments; D005337:Fibrin; D005340:Fibrinogen; D005176:Factor XIII; D010447:Peptide Hydrolases