title: The kinetic properties producing the perfunctory pH profiles of catalase-peroxidases.
abstract: Many structure-function relationship studies performed on the catalase-peroxidase enzymes are based on limited kinetic data. To provide a more substantive understanding of catalase-peroxidase function, we undertook a more exhaustive evaluation of catalase-peroxidase catalysis as a function of pH. Kinetic parameters across a broad pH range for the catalase and peroxidase activities of E. coli catalase peroxidase (KatG) were obtained, including the separate analysis of the oxidizing and reducing substrates of the peroxidase catalytic cycle. This investigation identified ABTS-dependent inhibition of peroxidase activity, particularly at low pH, unveiling that previously reported pH optima are clearly skewed. We show that turnover and efficiency of peroxidase activity increases with decreasing pH until the protein unfolds. The data also suggest that the catalase pH optimum is more complex than it is often assumed to be. The apparent optimum is in fact the intersection of the optimum for binding (7.00) and the optimum for activity (5.75). We also report the apparent pK(a)s for binding and catalysis of catalase activity as well as approximate values for certain peroxidatic and catalatic steps.
authors.affiliation: Department of Chemistry and Biochemistry, Auburn University, AL 36849, USA.
authors.forename: Robert L
authors.initials: RL
authors.lastname: Moore
authors.forename: Luke J
authors.initials: LJ
authors.lastname: Powell
authors.forename: Douglas C
authors.initials: DC
authors.lastname: Goodwin
mesh_terms: D002374:Catalase / Q000737:chemistry* / Q000378:metabolism*; D002384:Catalysis; D002942:Circular Dichroism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism*; D006418:Heme / Q000737:chemistry / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D017510:Protein Folding
chemical_list: D029968:Escherichia coli Proteins; D006418:Heme; D002374:Catalase; C051253:katG protein, E coli