Comparative sequence analysis of vitamin K-dependent coagulation factors

J Thromb Haemost. 2022 Dec;20(12):2837-2849. doi: 10.1111/jth.15897. Epub 2022 Oct 11.


Background: Prothrombin, protein C, and factors VII, IX, and X are vitamin K (VK)-dependent coagulation proteins that play an important role in the initiation, amplification, and subsequent attenuation of the coagulation response. Blood coagulation evolved in the common vertebrate ancestor as a specialization of the complement system and immune response, which in turn bear close evolutionary ties with developmental enzyme cascades. There is currently no comprehensive analysis of the evolutionary changes experienced by these coagulation proteins during the radiation of vertebrates and little is known about conservation of residues that are important for zymogen activation and catalysis.

Objectives: To characterize the conservation level of functionally important residues among VK-dependent coagulation proteins from different vertebrate lineages.

Methods: The conservation level of residues important for zymogen activation and catalysis was analyzed in >1600 primary sequences of VK-dependent proteins.

Results: Functionally important residues are most conserved in prothrombin and least conserved in protein C. Some of the most profound functional modifications in protein C occurred in the ancestor of bony fish when the basic residue in the activation site was replaced by an aromatic residue. Furthermore, during the radiation of placental mammals from marsupials, protein C acquired a cysteine-rich insert that introduced an additional disulfide in the EGF1 domain and evolved a proprotein convertase cleavage site in the activation peptide linker that also became significantly elongated.

Conclusions: Sequence variabilities at functionally important residues may lead to interspecies differences in the zymogen activation and catalytic properties of orthologous VK-dependent proteins.

Keywords: blood coagulation; evolution; protein C; prothrombin; vitamin K-dependent clotting factors.

MeSH terms

  • Animals
  • Blood Coagulation Factors / genetics
  • Blood Coagulation Factors / metabolism
  • Female
  • Mammals / metabolism
  • Placenta
  • Pregnancy
  • Protein C
  • Prothrombin* / metabolism
  • Sequence Analysis
  • Vitamin K* / metabolism


  • Vitamin K
  • Prothrombin
  • Protein C
  • Blood Coagulation Factors