Human plasma fibrinogen heterogeneity: evidence for an extended carboxyl-terminal sequence in a normal gamma chain...

Two types of normal human plasma fibrinogen--peak 1 and peak 2--are distinquishable by DEAE-cellulose gradient elution chromatography. The elution characteristics of peak 2 fibrinogen, which amounts to about 15% of the total, are attributable to the presence of a gamma chain variant, gamma', which is more negatively charged than gamma chains and makes up about half of all such chains in that peak [Mosesson M. W., Finlayson, J. S. & Umfleet, R. A. (1972), J. Biol. Chem. 247, 5223-5227]. Analyses of reduced S-carboxymethylated fibrin that had first been incubated in the presence of Factor XIIIa plus the fluorescent amine donor dansylcadaverine (DNScad) showed that the same amount of this compound could be incorporated covalently into either type of gamma chain. Furthermore, the DNScad-labeled COOH-terminal CNBr fragment (CNBr e) derived from the S-carboxymethylated gamma chain was smaller than the DNScad-labeled fragment (CNBr e') from the gamma' chain (Mr, 3200 and 4900) by about the same amount as the difference in size between the respective parent chains (Mr, 49,400 and 51,500). DNScad-CNBr e or DNScad-cNBR e' could be further cleaved by trypsin to yield a smaller fluorescent fragment corresponding to the penultimate tryptic gamma chain peptide containing the DNScad-glutamine acceptor and lysine donor crosslinking functions. The COOH-terminal amino acids of gamma and gamma' chains were valine and leucine, respectively. The rates of Factor XIIIa-catalyzed crosslinking of peak 1 and peak 2 fibrin were the same, but peak 1 fibrin gamma chains formed only one species of crosslinked dimer (gamma gamma) whereas peak 2 fibrin gamma chains yielded three (gamma gamma, gamma gamma', gamma'gamma'). We conclude that gamma' chains are functionally normal but have an extended COOH-terminal sequence accounting for their more negative charge and larger size relative to gamma chains.