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Examples of the kinetics of FRETS-VWF73 cleavage (1 μM) by 0.5 nM ADAMTS13 in the four-buffer system solution at pH 5.0 (▪) and 10.0 (•) and T = 25°C. For sake of clarity, the fluorescence signal at time = ∞ was set at 100% and corresponded to 1 μM cleaved FRETS-VWF73. The continuous lines were drawn according to a simple first order kinetics with the best-fit kcat/Km values equal to 5.2 ± 0.1 × 105 M−1 sec−1 and 2.03 ± 0.06 × 105 M−1 sec−1 for pH 5 and 10, respectively.
