Overall cleavage rate [(kcat/Km)s] and binding affinity (Kd) in cleavage reactions of tat37 with RNase P ribozymes.

Binding assays were carried out in buffer D (50 mM Tris, pH 7.5, 100 mM NH4Cl, 100 mM CaCl2, 3% glycerol, 0.1% xylene cyanol, 0.1% bromphenol blue), using a protocol modified from Pyle et al [38]. Single-turnover kinetic analyses to determine the values of (kcat/Km)s were carried out in buffer A (50 mM Tris-HCl, pH 7.5, 100 mM NH4Cl, 100 mM MgCl2) as described previously [36]. The values shown are the averages derived from five independent experiments.