Abstract
Antithrombin functions as the principal plasma protein inhibitor of most blood coagulation proteinases.1,2 The essential role of this inhibitor in regulating the activity of these proteinases in vivo is indicated from the well-established link between inherited or acquired deficiencies of antithrombin and the tendency to develop thrombotic disease. Antithrombin is a member of the serpin superfamily of protein proteinase inhibitors and its main target enzymes include the blood coagulation factors IXa, Xa and thrombin. This and other serpins are distinguished from other family members in that their reactions with target enzymes are greatly accelerated by the binding of heparin or heparan sulfate glycosaminoglycans. This property is chiefly responsible for the anticoagulant activity of heparin and has suggested a role for endogenous heparin and heparan sulfate in the regulation of blood coagulation proteinases by antithrombin. In this article, we will review our present understanding of the relationship between antithrombin structure and function based on currently available evidence. Our discussion will focus principally on two areas: 1) the mechanism by which antithrombin and other serpins inhibit their target proteinases; and 2) the molecular basis of heparin’s accelerating effect on antithrombin-proteinase reactions.
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References
I. Björk, S. T. Olson, and J. D. Shore, Molecular mechanisms of the accelerating effect of heparin on the reactions between antithrombin and clotting proteinases, in: “Heparin. Chemical and Biological Properties. Clinical Applications”, D. A. Lane and U. Lindahl, eds., Edward Arnold, London (1989).
S. T. Olson and I. Björk, Regulation of thrombin by antithrombin and heparin cofactor II, in: “Thrombin: Structure and function”, L. J. Berliner, ed., Plenum, New York City (1991).
I. Björk and W. W. Fish, Production in vitro and properties of a modified form of bovine antithrombin, cleaved at the active site by thrombin, J. Biol. Chem. 257:9487 (1982).
S. T. Olson, Heparin and ionic strength-dependent conversion of antithrombin III from an inhibitor to a substrate of a-thrombin, J. Biol. Chem. 260:10153 (1985).
H. Erdjument, D. A. Lane, M. Panico, V. DiMarzo, and H. R. Morris, Single amino acid substitutions in the reactive site of antithrombin leading to thrombosis. Congenital substitution of arginine 393 to cysteine in antithrombin Northwick Park and to histidine in antithrombin Glasgow. J. Biol. Chem. 263:5589 (1988).
M. C. Owen, C. H. Beresford, and R. W. Carrell, Antithrombin Glasgow, 393 Arg to His: a P1 reactive site variant with increased heparin affinity but no thrombin inhibitory activity, FEBS Lett. 231:317 (1988).
D. A. Lane, H. Erdjument, A. Flynn, V. DiMarzo, M. Panico, H. R. Morris, M. Greaves, G. Dolan, and F. E. Preston, Antithrombin Sheffield: amino acid substitution at the reactive site (Arg 393 to His) causing thrombosis, Brit. J. Haematol. 71:91 (1989).
H. Erdjument, D. A. Lane, M. Panico, V. DiMarzo, H. R. Morris, K. Bauer, and R. D. Rosenberg, Antithrombin Chicago, amino acid substitution of arginine 393 to histidine, Thromb. Res. 54:613 (1989).
H. Erdjument, D. A. Lane, H. Ireland, V. DiMarzo, M. Panico, H. R. Morris, A. Tripodi, and P. M. Manucci, Antithrombin Milano, single amino acid substitution at the reactive site, Arg 393 to Cys, Thromb. Hemostas. 60:471 (1988).
D. A. Lane, H. Erdjument, E. Thompson, M. Panico, V. DiMarzo, H. R. Morris, G. Leone, V. DeStefano, and S. L. Thein, A novel amino acid substitution in the reactive site of a congenital variant antithrombin. Antithrombin Pescara, Arg393 to Pro, caused by a CGT to CCT mutation, J. Biol. Chem. 264: 10200 (1989).
A. W. Stephens, B. S. Thalley, and C. H. W. Hirs, Antithrombin Ill-Denver, a reactive site variant, J. Biol. Chem. 262:1044 (1987).
A. W. Stephens, A. Siddiqui, and C. H. W. Hirs, Site-directed mutagenesis of the reactive center (serine 394) of antithrombin III, J. Biol. Chem. 263:15849 (1988).
S. T. Olson, R. Sheffer, A. W. Stephens, and C. H. W. Hirs, Molecular basis of the reduced reactivity of antithrombin-Denver with thrombin and factor Xa. Role of the P1’ residue, Thromb. Haemostas. 65:670 (1991).
S. C. Bock, Antithrombin III genetics, structure and function, in: “Recombinant Technology in Hemostasis and Thrombosis”, L. W. Hoyer and W. N. Drohan, eds., Plenum, New York City (1990).
P. Molho-Sabatier, M. Aiach, I. Gaillaird, J. N. Fiessinger, A. M. Fischer, G. Chadeuf, and E. Clause, Molecular characterization of antithrombin III (AT III) variants using polymerase chain reaction. Identification of the AT III Charleville as an Ala 384 →Pro mutation, J. Clin. Invest. 84:1236 (1989).
R. Caso, D. A. Lane, E. A. Thompson, R. J. Olds, S. L. Thein, M. Paqnico, I. Blench, H. R. Morris, J. M. Freyssinet, M. Aiach, F. Rodeghiero, and G. Finazzi, Antithrombin Vicenza, Ala 384 to Pro (GCA to CCA) mutation, transforming the inhibitor into a substrate, Brit. J. Haematol. 77:87 (1991).
R. Devraj-Kizuk, D. H. K. Chui, E. V. Prochownik, C. J. Carter, F. A. Ofosu, M. A. Blajchman, Antithrombin-III-Hamilton: A gene with a point mutation (guanine to adenine) in codon 382 causing impaired serine protease reactivity, Blood 72:1518 (1988).
N. J. Levy, N. Ramesh, M. Cicardi, R. A. Harrison, A. E. Davis, Type II hereditary angioneurotic edema that may result from a single nucleotide change in the codon for alanine-436 in the Cl inhibitor gene, Proc. Natl. Acad. Sci. U.S.A. 87:265 (1990).
K. Skriver, W. R. Wikoff, P. A. Patston, F. Tausk, M. Schapira, A. P. Kaplan, and S. C. Bock, Substrate properties of C1 Inhibitor Ma (alaninie 434→glutamic acid). Genetic and structural evidence suggesting that the P12-region contains critical determinants of serine protease inhibitor inhibitor/substrate status, J. Biol. Chem. 266:9216 (1991).
W. E. Holmes, H. R. Lijnen, L. Nelles, C. Kluft, H. K. Nieuwenhuis, D. C. Rijken, and D. Collen, α2-antiplasmin Enschede: alanine insertion and abolition of plasmin inhibitory activity, Science 238:209 (1987).
S. Asakura, H. Hirata, H. Okazaki, T. Hashimoto-Gotoh, and M. Matsuda, Hydrophobic residues 382–386 of antithrombin III, ala-ala-ala-ser-thr, serve as an epitope for an antibody which facilitates hydrolysis of the inhibitor by thrombin, J. Biol. Chem. 265:5135 (1990).
R. Huber and R. W. Carrell, Implications of the three-dimensional structure of α1-antitrypsin for structure and function of serpins, Biochemistry 28:8951 (1989).
L. Mourey, J. P. Samama, M. Delarue, J. Choay, J. C. Lormeau, M. Petitou, and D. Moras, Antithrombin III: structural and functional aspects, Biochimie 72:599 (1990).
R. A. Engh, H. T. Wright, and R. Huber, Modeling of the intact form of the α1-proteinase inhibitor, Protein Engng. 3:469 (1990).
P. E. Stein, A. G. W. Leslie, J. T. Finch, W. G. Turnell, P. J. McLaughlin, and R. W. Carrell, Crystal structure of ovalbumin as a model for the reactive centre of serpins, Nature 347:99 (1990).
A. J. Schulze, U. Baumann, S. Knof, E. Jaeger, R. Huber, and C.-B. Laurell, Structural transition of α1-antitrypsin by a peptide sequentially similar to β-strand s4A, Eur. J. Biochem. 194:51 (1990).
I. Björk, K. Ylinenjarvi, S. T. Olson, and P. E. Bock, Conversion of antithrombin from an inhibitor of thrombin to a substrate with reduced heparin affinity and enhanced conformational stability by binding of a tetradecapeptide corresponding to the P1 to P14 region of the putative reactive-bond loop of the inhibitor, J. Biol. Chem. in press (1991).
S. O. Brennan, J. Y. Borg, P. M. George, C. Soria, J. Soria, J. Caen, and R. W. Carrell, New carbohydrate site in mutant antithrombin (7 Ile→ Asn) with decreased heparin affinity, FEBS Lett. 237:118 (1988).
J. Y. Borg, S. O. Brennan, R. W. Carrell, P. George, D. J. Perry, and J. Shaw, Antithrombin Rouen-IV, 24 Arg→Cys. The amino terminal contribution to heparin binding, FEBS Lett. 266:163 (1990).
S. Gandrille, M. Aiach, D. A. Lane, D. Vidaud, P. Molho-Sabatier, R. Caso, P. deMoerloose, J. N. Fiessinger, and E. Clauser, Crucial role of Arg 129 in heparin binding site of antithrombin III: Identification of a novel mutation Arg 129 to Gln, J. Biol. Chem. 265:18997(1990).
P. Gettins and E. W. Wooten, On the domain structure of antithrombin III. Localization of the heparin-binding region using 1H NMR spectroscopy, Biochemistry 26:4403 (1987).
J. Y. Chang, Binding of heparin to human antithrombin III activates selective chemical modification at lysine 236. Lys-107, lys-125, and lys-136 are situated within the heparin-binding site of antithrombin III, J. Biol. Chem. 264:3111 (1989).
X. J. Sun and J. Y. Chang, Evidence that arginine-129 and arginine-145 are located within the heparin binding site of human antithrombin III, Biochemistry 29:8957 (1990).
X. J. Sun and J. Y. Chang, Heparin binding domain of human antithrombin III inferred from the sequential reduction of its three disulfide linkages. An efficient method for structural analysis of partially reduced proteins, J. Biol. Chem. 264:11288 (1989).
J. W. Smith, N. Dey, and D. J. Knauer, Heparin binding domain of antithrombin III: Characterization using a synthetic peptide directed polyclonal antibody, Biochemistry 29:8950 (1990).
S. T. Olson, I. Björk, P. A. Craig, J. D. Shore, and J. Choay, Role of the high-affinity pentasaccharide in heparin acceleration of antithrombin III inhibition of thrombin and factor Xa, Thromb. Haemostas. 58:8 (1987).
R. D. Rosenberg and P. S. Damus, The purification and mechanism of action of human antithrombin-heparin cofactor, J. Biol. Chem. 248:6490 (1973).
C. H. Beresford and M. C. Owen, Minireview. Antithrombin III, Int. J. Biochem. 22:121 (1990).
S. T. Olson and I. Björk, Predominant contribution of surface approximation to the mechanism of heparin acceleration of the antithrombin-thrombin reaction. Elucidation from salt concentration effects, J. Biol. Chem. 266:6353 (1991).
S. T. Olson, High molecular weight-kininogen enhancement of the heparin-accelerated rate of plasma kallikrein inactivation by antithrombin III, J. Cell Biol. 107:827a (1989).
S. T. Olson and J. D. Shore, High molecular weight-kininogen and heparin acceleration of factor XIa inactivation by plasma proteinase inhibitors, Thromb. Haemostas. 62:381 (1989).
I. Björk, S. T. Olson, R. G. Sheffer, and J. D. Shore, Binding of heparin to human high molecular weight kininogen, Biochemistry 28:1213 (1989).
S. T. Olson and J. Choay, Mechanism of high molecular weight-kininogen stimulation of the heparin-accelerated antithrombin/kallikrein reaction, Thromb. Haemostas. 62:326 (1989).
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Olson, S.T., Björk, I. (1992). Role of Protein Conformational Changes, Surface Approximation and Protein Cofactors in Heparin-Accelerated Antithrombin-Proteinase Reactions. In: Lane, D.A., Björk, I., Lindahl, U. (eds) Heparin and Related Polysaccharides. Advances in Experimental Medicine and Biology, vol 313. Springer, Boston, MA. https://doi.org/10.1007/978-1-4899-2444-5_16
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DOI: https://doi.org/10.1007/978-1-4899-2444-5_16
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