title: Conservation of human fibrinogen conformation after cleavage of the B beta chain NH2 terminus.
abstract: Human fibrinogen exposed to protease III from Crotalus atrox venom is cleaved near the NH2 terminus of the B beta chain yielding a species of Mr 325,000 (Fg325) with impaired thrombin clottability. The derivative was compared with intact fibrinogen in a number of ways to determine whether the functional defect resulted from a conformational change or from the loss of a polymerization site. NH2-terminal amino acid sequencing of isolated A alpha, B beta, and gamma chains showed that Fg325 contained intact A alpha and gamma chains, but differed from fibrinogen by the absence of the first 42 residues of the B beta chain. Fibrinopeptide A was present and was cleaved at the same rate in both fibrinogen and Fg325. The rate and extent of A alpha and gamma cross-linking by factor XIIIa was also indistinguishable. In contrast, the thrombin-catalyzed coagulation of Fg325 was 46% less in extent and 180-fold slower than observed for intact fibrinogen. A conformational comparison of Fg325 and fibrinogen was made using immunochemical and spectroscopic approaches. Antisera specific for different regions of the fibrinogen molecule were used to characterize the epitopes in Fg325. The only significant differences were found in the NH2-terminal region of the B beta chain, probed with antiserum to B beta 1-118. The conformational similarity of Fg325 and fibrinogen was confirmed by the identity of both near and far UV CD spectra of the two proteins. Structural, functional, and immunochemical results imply that cleavage of 42 NH2-terminal residues from the B beta chain is not accompanied by a measurable conformational change. The residues of this B beta chain segment, which are evidently located on the surface of the molecule, in conjunction with the NH2-terminal part of the A alpha chain appear to play an important role in the expression of a fibrin polymerization site.
authors.forename: B V
authors.initials: BV
authors.lastname: Pandya
authors.forename: C S
authors.initials: CS
authors.lastname: Cierniewski
authors.forename: A Z
authors.initials: AZ
authors.lastname: Budzynski
mesh_terms: D000595:Amino Acid Sequence; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis; D005338:Fibrin Fibrinogen Degradation Products / Q000032:analysis* / Q000494:pharmacology; D005340:Fibrinogen / Q000378:metabolism*; D005344:Fibrinopeptide A / Q000032:analysis; D005345:Fibrinopeptide B / Q000032:analysis; D006801:Humans; D007700:Kinetics; D011487:Protein Conformation; D012965:Sodium Chloride / Q000494:pharmacology
chemical_list: D000939:Epitopes; D005338:Fibrin Fibrinogen Degradation Products; D005344:Fibrinopeptide A; D005345:Fibrinopeptide B; D012965:Sodium Chloride; D005340:Fibrinogen