Reaction of antithrombin III with thrombin bound to the vascular endothelium. Analysis in a recirculating perfused rabbit heart preparation.
A recirculating perfused rabbit heart preparation is used to study the reaction of antithrombin III (ATIII) with thrombin bound to the surface of the microvascular endothelium. Addition of ATIII to the system after thrombin is equilibrated with its binding sites results in inhibition of the enzyme as measured by disappearance of thrombin enzymatic activity from the circulation or by appearance of 125I-thrombin-ATIII complexes. The rate of inhibition of thrombin as reflected by either method is independent of the bound state of thrombin. Comparable results are obtained with ATIII modified at a single tryptophan residue. This modification does not alter the reaction rate of ATIII with thrombin but abolishes the capacity of heparin or heparan sulfate to enhance the reaction rate. From the kinetics and structural studies and the fit of the kinetics to a theoretical model relating binding equilibrium to thrombin inhibition, it is concluded that glycosaminoglycans are not involved in the reversible, high affinity, high capacity binding of thrombin to the vascular endothelium.[1]References
- Reaction of antithrombin III with thrombin bound to the vascular endothelium. Analysis in a recirculating perfused rabbit heart preparation. Lollar, P., MacIntosh, S.C., Owen, W.G. J. Biol. Chem. (1984) [Pubmed]
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