Thermal denaturation of cytochrome c peroxidase: pH dependence.

Upon heating cytochrome c peroxidase (ferrocytochrome c: hydrogen-peroxide oxidoreductase, EC 1.11.1.5) at pH 4 and 5, the enzyme precipitates at 41 degrees C and 51 degrees C, respectively. Incubating the enzyme at lower temperatures causes a slow dissociation of the heme from the protein. The heme precipitates, while the apoprotein remains soluble. Between pH 6 and 8, the native enzyme is converted to a low-spin ferric form upon heating. The Soret maximum shifts from 408 to 414 nm. The midpoint of this transition is pH-dependent, with a value of 46 degrees C at pH 6 decreasing to 29 degrees C at pH 8. At high temperatures the 414 nm form is converted to a species which has a 'free heme' spectrum with low absorptivity and Soret maximum at 390 nm. The midpoint temperature of this latter transition is 62 degrees C and 57 degrees C at pH 7 and 8, respectively.