Mechanism of interaction between tissue factor pethway inhibitor and heparin

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Surface plasmon resonance was developed to analyze the heparin-protein interaction using as ligands a series of heparin derivatives regioselectively desulfated by chemical methods, particularly in order to evaluate the effect of each sulfate group of heparin. The method for immobilizing heparin on the cuvette of the surface plasmon resonance equipment was optimized in order to obtain the high response required for accurate measurement. The best result was achieved when the amino group introduced at the reducing end of heparin was coupled with carboxymethyl dextran on the surface of the cuvette using glycolchitosan as a multi-valent linker. The established system appeared to describe well the interactions of heparin with such proteins as acidic and basic fibroblast growth factors and tissue factor pathway inhibitor.Using the combination of surface plasmon resonance and regioselectively heparins, the role of sulfate groups of heparin in the interaction with heparin was revealed. The results obtained indicated that all sulfate groups, 2-O, 2-N, and 6-O, contribute in the interaction of both molecule, and the contribution of 6-O-sulfate group is the largest.