The heparin-catalysed inhibition of human factor XIa by antithrombin III is dependent on the heparin type.
The effect of various well-characterized heparin preparations on the inactivation of human Factor XIa by human antithrombin III was studied. The heparin preparations used were unfractionated heparin and four heparin fractions obtained after anion-exchange chromatography. Inactivation of Factor XIa was monitored with S2366 as chromogenic substrate and followed pseudo-first-order reaction kinetics under all reaction conditions tested. Enhancement of the rate of inhibition of Factor XIa in the presence of unfractionated heparin correlated to the binding of antithrombin III to heparin. From the kinetic data a binding constant of 0.1 microM was inferred. The maximum rate enhancement, achieved at saturating heparin concentrations, was 30-fold. The rate enhancement achieved in the presence of each of the heparin fractions could also be correlated to the binding of antithrombin III to the heparin. The binding constant inferred from the kinetic data varied from 0.10 to 0.28 microM and the number of binding sites for antithrombin III varied from 0.06 to 0.74 site per heparin molecule. The maximum rate enhancements, achieved at saturating heparin concentrations, were strongly dependent on the type of heparin used and varied from 7-fold for fraction A to 41-fold for fraction D. Therefore, although the stimulation of Factor XIa inactivation by antithrombin III could be quantitatively correlated to the binding of antithrombin III to heparin, the heparin-catalysed inhibition of Factor XIa is dependent not only upon the degree of binding of antithrombin III to heparin but also upon the type of heparin to which antithrombin III is bound.
Soons
H
H
Department of Biochemistry, University of Limburg, Maastricht, The Netherlands.
Tans
G
G
Hemker
H C
HC
D000990:Antithrombin III / Q000378:metabolism*; D002384:Catalysis; D005591:Chemical Fractionation; D005172:Factor XI / Q000037:antagonists & inhibitors*; D015945:Factor XIa; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D007700:Kinetics
D000990:Antithrombin III; D006493:Heparin; D005172:Factor XI; D015945:Factor XIa