Biochemical characterization of the bacterial peroxidase from the human pathogen Neisseria gonorrhoeae

Cláudia S. Nóbrega, Mariana Raposo, Gonzalez Van Driessche, Bart Devreese, Sofia R. Pauleta

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Neisseria gonorrhoeae is an obligate human pathogen that expresses an array of molecular systems to detoxify reactive oxygen species as defense mechanisms during colonization and infection. One of these is the bacterial peroxidase that reduces H2O2 to water in its periplasm. The soluble form of this enzyme was heterologously expressed in E. coli in the holo-form binding two c-types hemes, a high-potential E heme and a low-potential P heme, with redox potentials of (+ 310 mV) and (− 190 mV/− 300 mV), respectively in the presence of calcium ions, at pH 7.5. Visible and EPR spectroscopic analysis together with activity assays indicate the presence of a calcium dependent reductive activation mechanism in thgonorrhoeaeNeisseria gonorrhoeae bacterial peroxidase, in which P heme is bis-His coordinated low-spin in the fully oxidized state of the enzyme, and becomes penta-coordinated high-spin upon reduction of E heme in the presence of calcium ions. The activated enzyme has a high affinity for H2O2 (KM of 4 ± 1 μM), with maximum activity being attained at pH 7.0 and 37 °C, with the rate-limiting step in the catalytic cycle being the electron transfer between the two hemes. In this enzyme, dimer formation is not promoted at high ionic strength, thus differing from the classical bacterial peroxidases. These results contribute to the understanding of the involvement of Neisseria gonorrhoeae bacterial peroxidase has a first line defense mechanism against exogenously produced hydrogen peroxide in the host environment.

Original languageEnglish
Pages (from-to)108-119
Number of pages12
JournalJournal of Inorganic Biochemistry
Volume171
DOIs
Publication statusPublished - 1 Jun 2017

Keywords

  • Bacterial peroxidase
  • Heme enzyme
  • Kinetics
  • Neisseria gonorrhoeae
  • Reactive oxygen species

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