Skip to Content

Human Heparin Cofactor II


Inhibition of Thrombin by Heparin Cofactor II
Thrombin (IIa), heparin (HEP) and heparin cofactor II (HCII) interact to form a ternary complex via a random order bireactant mechanism. Heparin subsequently dissociates from the complex as thrombin forms a covalent complex with the heparin cofactor II.

  • Price $365.00/100µg ($331.00/min. 5)
    Size 100µg
    Formulation 50% (vol/vol) Glycerol/H2O
    Molecular Weight
    Storage -20°C
    Purity >95% by SDS-PAGE
    Compound
    Assay Thrombin inhibition
    Shelf Life (properly stored) 12 months
    Chemical Formula
Gel Novex 4-12% Bis-Tris
Load Human Heparin Cofactor II, 1 µg per lane
Buffer MOPS
Standard SeeBluePlus 2; Myosin (191 kDa), Phosphorylase B (97 kDa), BSA (64 kDa), Glutamic Dehydrogenase (51 kDa), Alcohol Dehydrogenase (39 kDa), Carbonic Anhydrase (28 kDa), Myoglobin Red (19 kDa), Lysozyme (14 kDa)

Heparin cofactor II (HCII) (heparin cofactor A, antithrombin BM, dermatan sulfate cofactor, human leuserpin-2) is a single chain glycoprotein (Mr=65,600, 10% carbohydrate) member of the serine protease inhibitor (serpin) family (1-3). Amino acid sequence analysis of heparin cofactor II reveals 3 potential N-glycosylation sites and 2 sulfated tyrosines located in two internal repeats of 7 residues located near the NH2-terminal (4-6). The plasma concentration is estimated to be 90 µg/ml (2).

In the coagulation cascade, heparin cofactor II inhibits thrombin by formation of a bimolecular complex in the presence (k2=4.5x108 M-1min-1) and absence (k2=5.0x105 M-1min-1) of heparin (2, 7-9). This complex is stable to denaturants and essentially irreversible. In addition to heparin, a wide variety of polyanionic compounds accelerate inhibition of thrombin by HCII (3). Noteworthy among these compounds is dermatan sulfate, which accelerates inhibition of thrombin by HCII, but not antithrombin III (1,6) This feature has been utilized to develop a plasma based assay specific for heparin cofactor II. Although HCII shows only ~25% sequence homology to other serpins, the reactive site peptide (COOH-terminal 36 amino acids) released on inhibition of thrombin is 53% homologous to that released from antithrombin III (AT III) by thrombin (4,9). The scissile bond in HCII is Leu-Ser which is unusual for an arginine specific protease inhibitor (4,9). In contrast to AT III, the only coagulation protease inhibited by HCII is thrombin, Other proteases inhibited by HCII include α-chymotrypsin, neutrophil cathepsin G and streptomyces griseus protease B (3). The major physiologic role of heparin cofactor II may remain to be discovered.

Heparin cofactor II is prepared from fresh frozen plasma by a modification of the procedure of Griffith, et al., (9). Purity is assessed on SDS PAGE and activity is determined based on thrombin inhibition in the presence of dermatan sulfate. HCII is supplied in 50% (vol/vol) glycerol/H20 for storage at -20°C.

Localization Plasma
Plasma concentration 90 µg/ml (2)
Mode of action serine protease inhibitor; inhibits thrombin, a-chymotrypsin, neutrophil cathepsin G, streptomyces griseus protease B
Molecular weight 65,600 (2)
Extinction coefficient
E
1 %
1 c m, 280 nm
= 5.93 (5)
Isoelectric point 4.95-5.15 (2)
Structure single chain glycoprotein, 3 potential N-glycosylation sites, two 7-amino acid residue repeats, reactive site sequence: TVTTVGFMPL-STQVRFTVDR (4)
Percent carbohydrate 9.9% (2)
Post-translational modifications 3 sulfated tyrosines
  1. Bringinshaw, G.W. and Shanberge, J.M., Arch. Biochem. Biophys., 161, 683 (1974).
  2. Tollefsen, D.M., et al., J. Biol. Chem., 257, 2162 (1982).
  3. Pratt, C.W., et al., Ann. N.Y. Acad. Sci., 556, 104 (1989).
  4. Ragg, H., Nucleic Acids Res., 14, 1073 (1986).
  5. Church, F.C., et al., Arch. Biochem. Biophys., 259, 331 (1987).
  6. Hortin, G., et al., J. Biol. Chem., 261, 15827 (1986).
  7. Church, F.C., et al., Proc. Natl. Acad. Sci. USA, 82, 6431 (1985).
  8. Tollefsen, D.M. and Blank, M.K., J. Clin. Invest., 68, 589 (1981).
  9. Griffith, M.J., et al., J. Biol. Chem., 260, 2218 (1985).
  10. Tollefsen, D.M., et al., J. Biol. Chem., 258, 6713 (1983).
  11. Griffith, M.J. and Marbet, G.A., Biochem. Biophys. Res. Comm., 112, 663 (1983).
  12. Abildgaard, U. and Larsen, M.L., Thromb. Res., 35, 257 (1984).
  13. Griffith, M.J., et al., Biochemistry, 24, 6777 (1985).
  14. Parker, K. and Tollefsen, D.M., J. Biol. Chem., 260, 3501 (1985).
  1. Monteiro, R., et al., Biochem. J. (2005) 387, 871–877. (Prothrombin Activation)
  2. Reis, C., et al., Biochem. J. (2006) 398, 295–302 (Prothrombin Activation)