Inhibition of trypsin by heparin and dalteparin, a low molecular weight heparin

The interaction between trypsin, a prototype S1 serine protease, with heparin and its low molecular weight derivative dalteparin were investigated. Direct inhibition of the proteolytic activity of trypsin by heparin and dalteparin, used in concentrations typical for their clinical application, was detected. The half-maximum inhibition of the trypsin activity was achieved at 15.25±1.22 μg/mL for heparin and was estimated to be at 58.47±15.20 μg/mL for dalteparin. Kinetic analyses showed that heparin and its low molecular weight derivative dalteparin inhibited trypsin by occupation of an exosite, producing noncompetitive and mixed inhibition, respectively. Heparin as a noncompetitive inhibitor with constant of inhibition Ki1,2 = 0.151±0.019 μM and dalteparin with Ki1 = 0.202±0.030 μM and Ki2 = 0.463±0.069 μM in mixed inhibition both represent moderate inhibitors of serine protease trypsin. The obtained constants of inhibition indicate that under the clinically applied concentrations of heparin and dalteparin, trypsins and their homolog S1 serine proteases could be directly inhibited, influencing the delicate control of proteolytic reactions in