Figure 5: Temperature dependent proteolysis of AGT variants by thermolysin. (a) thermal scans of AGT-LRM (10 μM in protein subunit) in the presence of different thermolysin concentrations (0–0.4 mg/mL). The high temperature transition ( ~ 90°C) corresponds to thermolysin denaturation, which is thermostable. (b) Dependence of downshift of thermal transitions of AGT variants by thermolysin. (c) Dependence of the apparent proteolysis rate constants on thermolysin concentration at 44°C. The slope of these plots provides the second-order rate constants (plotted in (d)); (d) Arrhenius plots for the second-order rate constants for proteolysis. The vertical dashed line indicates 37°C. Experiments were performed in Hepes 20 mM NaCl 200 mM CaCl2 10 mM.