Biochemical Characterization of the Copper Nitrite Reductase from Neisseria gonorrhoeae

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Abstract

The copper-containing nitrite reductase from Neisseria gonorrhoeae has been shown to play a critical role in the infection mechanism of this microorganism by producing NO and abolishing epithelial exfoliation. This enzyme is a trimer with a type 1 copper center per subunit and a type 2 copper center in the subunits interface, with the latter being the catalytic site. The two centers were characterized for the first time by EPR and CD spectroscopy, showing that the type 1 copper center has a high rhombicity due to its lower symmetry and more tetragonal structure, while the type 2 copper center has the usual properties, but with a smaller hyperfine coupling constant (A// = 10.5 mT). The thermostability of the enzyme was analyzed by differential scanning calorimetry, which shows a single endothermic transition in the thermogram, with a maximum at 94 °C, while the CD spectra in the visible region indicate the presence of the type 1 copper center up to 80 °C. The reoxidation of the N. gonorrhoeae copper-containing nitrite reductase in the presence of nitrite were analyzed by visible spectroscopy and showed a pH dependence, being higher at pH 5.5–6.0. The high thermostability of this enzyme may be important to maintaining a high activity in the extracellular space and to making it less susceptible to denaturation and proteolysis, contributing to the proliferation of N. gonorrhoeae.
Original languageEnglish
Article number1215
Number of pages15
JournalBiomolecules
Volume13
Issue number8
DOIs
Publication statusPublished - 4 Aug 2023

Keywords

  • AniA
  • copper enzyme
  • copper nitrite reductase
  • hyperthermostable
  • Neisseria
  • nitrite reduction
  • spectroscopy
  • T1 copper center
  • T2 copper center
  • thermostability

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