1. The reversible proton dissociation equilibria of peroxidase, apoperoxidase and haem-recombined apoperoxidase have been explored in 150mm-potassium chloride at 20°C at pH3–11.5. 2. Complementary heat measurements have been made of the classes of titratable groups to determine their intrinsic ΔH dissociation. 3. These curves are interpreted as showing that there are two histidine residues capable of titration in peroxidase whereas there are three such in apoperoxidase. 4. Concomitant spectroscopic investigations indicate profound differences in the tyrosine ionizations in the two proteins. In peroxidase one group only of the five residues ionizes up to pH11.5. In apoperoxidase four residues are titratable. 5. Spectroscopic titration in 6m-guanidinium chloride and 150mm-potassium chloride reveal one tyrosine residue fewer in peroxidase than in apoperoxidase. 6. These findings are discussed in terms of the ‘side chain’ groups responsible for binding the haem group in peroxidase. A proximal imidazole group seems probable as is also the involvement of a distally placed tyrosine. 7. The differences between apo- and holo-peroxidase are stressed, particularly in respect of abnormal carboxyl group titration in the former.
Research Article|September 01 1971
The hydrogen ion equilibria of horseradish peroxidase and apoperoxidase
Biochem J (1971) 124 (3): 605-614.
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C Phelps, L Forlani, E Antonini; The hydrogen ion equilibria of horseradish peroxidase and apoperoxidase. Biochem J 1 September 1971; 124 (3): 605–614. doi: https://doi.org/10.1042/bj1240605
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