The Catalase Activity of Catalase-Peroxidases Is Modulated by Changes in the pK a of the Distal Histidine

Biochemistry. 2017 May 2;56(17):2271-2281. doi: 10.1021/acs.biochem.6b01276. Epub 2017 Apr 20.

Abstract

The unusual Met-Tyr-Trp adduct composed of cross-linked side chains along with an associated mobile Arg is essential for catalase activity in catalase-peroxidases. In addition, acidic residues in the entrance channel, in particular an Asp and a Glu ∼7 and ∼15 Å, respectively, from the heme, significantly enhance catalase activity. The mechanism by which these channel carboxylates influence catalase activity is the focus of this work. Seventeen new variants with fewer and additional acidic residues have been constructed and characterized structurally and for enzymatic activity, revealing that their effect on activity is roughly inversely proportional to their distance from the heme and adduct, suggesting that the electrostatic potential of the heme cavity may be affected. A discrete group of protonable residues are contained within a 15 Å sphere surrounding the heme iron, and a computational analysis reveals that the pKa of the distal His112, alone, is modulated within the pH range of catalase activity by the remote acidic residues in a pattern consistent with its protonated form having a key role in the catalase reaction cycle. The electrostatic potential also impacts the catalatic reaction through its influence on the charged status of the Met-Tyr-Trp adduct.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Biocatalysis
  • Burkholderia pseudomallei / enzymology*
  • Catalase / chemistry
  • Catalase / genetics
  • Catalase / metabolism*
  • Catalytic Domain
  • Computational Biology
  • Crystallography, X-Ray
  • Hemeproteins / chemistry
  • Hemeproteins / genetics
  • Hemeproteins / metabolism*
  • Histidine / chemistry*
  • Hydrogen-Ion Concentration
  • Models, Molecular*
  • Mutation
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / metabolism
  • Peroxidases / chemistry
  • Peroxidases / genetics
  • Peroxidases / metabolism*
  • Protein Conformation
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Static Electricity
  • Titrimetry

Substances

  • Bacterial Proteins
  • Hemeproteins
  • Peptide Fragments
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Histidine
  • Peroxidases
  • Catalase