Binding of different forms of thrombin to HCII 1-75 immobilized on microtiter plate wells. Binding of α-thrombin (α-IIa, solid squares) or γT-thrombin (γT-IIa, ▾) was measured as the optical density (OD) at 490 nm as described in Materials and Methods, and graphed relative to the mean OD of binding reactions carried out with 100 nM α-thrombin, taken as 100%, in Panel A. The mean ± the SD of 3 independent experiments is shown. Panel B shows the results of experiments similar to those shown in Panel A, but carried out with 12.5 nM α-thrombin (α-IIa), S195A-thrombin (S195A-IIa), FPR-ck-thrombin (FPR-ck-IIa) or γT-thrombin (γT-IIa); results were normalized to the mean optical density (OD) at 490 nm observed for α-thrombin binding. Each bar represents the mean ± the SD of 3 independent experiments; * indicates p<0.05 by non-parametric ANOVA with Dunn’s post-test versus α-thrombin binding.
Boyle et al. BMC Biochemistry 2013 14:6 doi:10.1186/1471-2091-14-6