Biosynthesis of plasma factor XIII: evidence for transcription and translation in hepatoma cells

Abstract

Factor XIIIa belongs to a family of ubiquitous transglutaminases, which catalyze formation of covalent bonds between the ϵ-amino group of specific lysines and the γ-carboxyl group of glutamines. Factor XIII is synthesized as a zymogen and after activation, it participates in both the coagulation and fibrinolytic mechanisms. Most transglutaminases are intracellular, but factor XIII is both intracellular and extracellular. The biosynthesis of extracellular (plasma) factor XIII, with the structure of a noncovalent heterotetramer, A 2B 2, is complex. Here, evidence is presented from PCR analysis and Northern blotting that mRNAs for both A and B subunits are present in the liver. The distribution of mRNA, specific for factor XIII subunits, in various human tissues was also analyzed. Among the tissues examined, the only signal for B subunit was found in the liver. For subunit A, the signal was observed in placenta, liver, kidney, lung, skeletal muscle and heart with varying intensities; in brain or pancreas there was no signal. With an immunoperoxidase method, factor XIII A subunit was identified in the PLC/PRF/5 cell line. By ELISA and reverse immunoblotting, with antibodies specific for the A-B complex, it was also shown that these cells produce and secrete factor XIII. From all of these results, we conclude that the liver is a source of plasma factor XIII, and that the complex A 2B 2 is secreted from these cells.