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Thromb Haemost 1994; 71(06): 778-782
DOI: 10.1055/s-0038-1642522
Review Article
Schattauer GmbH Stuttgart

Molecular Cloning and Expression of Rabbit Heparin Cofactor II: A Plasma Thrombin Inhibitor Highly Conserved between Species

William P Sheffield
1   The Canadian Red Cross Society Blood Services McMaster University, Hamilton, Ontario, Canada
2   The Department of Pathology, McMaster University, Hamilton, Ontario, Canada
,
Philip D Schuyler
2   The Department of Pathology, McMaster University, Hamilton, Ontario, Canada
,
Morris A Blajchman
1   The Canadian Red Cross Society Blood Services McMaster University, Hamilton, Ontario, Canada
2   The Department of Pathology, McMaster University, Hamilton, Ontario, Canada
› Author Affiliations
Further Information

Publication History

Received: 21 December 1993

Accepted after revision: 05 February 1994

Publication Date:
09 July 2018 (online)

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Summary

Heparin cofactor II (HCII), a circulating plasma protein that inhibits thrombin, is a member of the serine proteinase (serpin) family of proteins. The extent to which HCII structure is conserved actross species lines was investigated, by obtaining cDNA clones encoding rabbit HCII. Overlapping clones corresponding to rabbit HCII were obtained by the combined use of hybridization screening of a rabbit liver cDNA library, and by rapid amplification of cDNA ends (RACE). The consensus sequence obtained spans 2178 nucleotides, and is comprised of a 5' untranslated region of 77 nucleotides, an open reading frame of 1440 nucleotides, and 3' untranslated region of 661 nucleotides that concludes with a poly A tract. The open reading frame is subdivided into a secretory signal sequence of 19 amino acids, and a mature protein of 461 amino acids. Within the region comprising the mature protein, 87% of the amino acid residues are identical to those seen in human HCII. Expression of an appropriately modified form of the rabbit HCII clone in an in vitro reticulocyte expression system yielded two major polypeptides, of 60 and 56 kD respectively, both of which were able to form SDS-stable complexes with human α-thrombin, in a reaction accelerated by dermatan sulphate. The remarkable degree of homology observed between rabbit HCII and its human conterpart, indicating a high degree of conservation of structure through evolution, suggests an important function of HCII in hemostatis.

 

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