Proteolytic activation of protein C from bovine plasma

Abstract

Protein C is a vitamin K dependent protein present in bovine plasma (Stenflo, J. (1976), J. Biol. Chem. 251, 355). It is a glycoprotein (mol wt approximately 62 000) composed of a heavy chain (mol wt 41 000) and a light chain (mol wt 21 000). The heavy chain has an amino-terminal sequence of Asp-Thr-Asn-Gln and contains nearly three-fourths of the carbohydrate. The light chain has an amino-terminal sequence of Ala-Asn-Ser-Phe. Incubation of protein C with either factor X activator from Russell's viper venom or trypsin resulted in the cleavage of an Arg-Ile bond between residues 14 and 15 of the heavy chain. Concomitant with this cleavage was the formation of a serine enzyme which was inhibited by diisopropyl phosphorofluoridate. Liberation of the tetradecapeptide decreased the molecular weight of the heavy chain from about 41 000 to 39 000 and resulted in the formation of a new amino-terminal sequence of Ile-Val-Asp-Gly in the heavy chain. No change in the molecular weight of the light chain was observed during the activation reaction. These results indicate that protein C, like the four vitamin K dependent coagulation proteins, exists in plasma in a precursor form and is converted to a serine protease by hydrolysis of a specific Arg-Ile peptide bond. The biological substrate for the enzymatic form of protein C and the physiological mechanism whereby protein C is converted to a serine enzyme are not known.