Measurement of the affinities of heparins, naturally occurring glycosaminoglycans, and other sulfated polymers for antithrombin III and thrombin

Anal Biochem. 1988 Oct;174(1):177-86. doi: 10.1016/0003-2697(88)90533-7.


Heparin, other glycosaminoglycans, and synthetic sulfated polymers have antithrombotic and anticoagulant activities, which may be mediated through a range of interactions with different proteins. A simple, quantitative method has been developed for assessing the affinity of interaction between sulfated polymers and proteins in the liquid phase. This has been used to compare the binding of a range of glycosaminoglycans and other sulfated polymers to antithrombin III and thrombin, a major inhibitor of and a central protease in the coagulation system, respectively. The results are consistent with the binding of naturally occurring glycosaminoglycans to antithrombin III solely through the well-defined antithrombin III-binding pentasaccharide found in heparin, the apparent affinity of a preparation depending upon its content of this pentasaccharide. Highly sulfated synthetic polymers will, however, bind antithrombin III by a second mechanism. The affinity of heparin for thrombin decreased with decreasing molecular weight. However, results obtained with heparan sulfate preparations did not indicate any clear relationship between either molecular weight or sulfate content and thrombin binding, but suggested that there may be an oligosaccharide sequence containing N-sulfate residues which confers high affinity for thrombin. In addition, some of the synthetic sulfated polymers bound thrombin with very high affinity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antithrombin III / metabolism*
  • Binding Sites
  • Binding, Competitive
  • Glycosaminoglycans / metabolism*
  • Heparin / metabolism*
  • Humans
  • In Vitro Techniques
  • Kinetics
  • Protein Binding
  • Sulfuric Acids / metabolism
  • Thrombin / metabolism*


  • Glycosaminoglycans
  • Sulfuric Acids
  • Antithrombin III
  • Heparin
  • Thrombin