Download PDF
Thromb Haemost 1993; 69(05): 422-429
DOI: 10.1055/s-0038-1651627
Original Article
Coagulation
Schattauer GmbH Stuttgart

Self-Association of Antithrombin III Relates to Multimer Formation of Thrombin-Antithrombin III Complexes

Klaus T Preissner
The Haemostasis Research Unit, Kerckhoff-Klinik, MPG, Bad Nauheim, Germany
› Author Affiliations
Further Information

Publication History

Received 30 June 1992

Accepted after revision 04 January 1993

Publication Date:
25 July 2018 (online)

Also available at
  PDF Download Buy Article Permissions and Reprints
Zoom Image

Summary

During the reaction of antithrombin III (AT III) with target proteases the inhibitor serves as pseudo-substrate and undergoes profound conformational changes, becomes incorporated into a covalent stoichiometric enzyme-inhibitor complex which is, in contrast to native AT III, recognized by monoclonal antibody 4C9. In the absence of the target enzyme thrombin, incubation of AT III with 1–2 M guanidine, 0.6% deoxycholate, heating to 56° C, or buffer at pH 4 resulted in inactivation of the inhibitor with concomitant exposure of the epitope for 4C9 and formation of AT III multimers (from 3.9S to 7.1–7.4 S). Loss of activity, formation of multimers and exposure of neoepitope(s) of AT III occurred in a concerted fashion and followed second order kinetics with an activation energy of E a = 31 kcal/mol. AT III-multimerization induced by treatment with 1 M guanidine (mainly AT III-tetramers with M r of 250,000) and formation of the binary AT III-thrombin complex revealed similar self-association patterns as judged by gel electrophoresis under non-denaturing conditions. In the presence of heparin, even higher multimers of AT III-thrombin complexes were noted. Moreover, self-association products of the ternary vitronectin-thrombin-AT III complex, which is the ultimate reaction product following thrombin inhibition in the circulation, could be recognized and quantitated due to exposure of the 4C9 epitope on AT III, indicating that AT III exists in multimeric forms within binary and ternary complexes. It is proposed that the ability of these complexes to form high M r association products is at least in part mediated by the propensity of AT III to multimerize and that multimeric forms of AT III may be involved during clearance of AT III-enzyme complexes.