Protonation state of the Cu4S2 Cu-Z site in nitrous oxide reductase: redox dependence and insight into reactivity

Esther M. Johnston, Simone Dell'Acqua, Sofia R. Pauleta, Isabel Moura, Edward I. Solomon

Research output: Contribution to journalArticle

Abstract

Spectroscopic and computational methods have been used to determine the protonation state of the edge sulfur ligand in the Cu4S2 CuZ form of the active site of nitrous oxide reductase (N2OR) in its 3Cu(I)Cu(II) (1-hole) and 2Cu(I)2Cu(II) (2-hole) redox states. The EPR, absorption, and MCD spectra of 1-hole Cu-Z indicate that the unpaired spin in this site is evenly delocalized over Cu-I, Cu-II, and Cu-IV. 1-hole Cu-Z is shown to have a mu(2)-thiolate edge ligand from the observation of S-H bending modes in the resonance Raman spectrum at 450 and 492 cm(-1) that have significant deuterium isotope shifts (-137 cm(-1)) and are not perturbed up to pH 10. 2-hole CuZ is characterized with absorption and resonance Raman spectroscopies as having two Cu-S stretching vibrations that profile differently. DFT models of the 1-hole and 2-hole Cu-Z sites are correlated to these spectroscopic features to determine that 2-hole Cu-Z has a mu(2)-sulfide edge ligand at neutral pH. The slow two electron (+1 proton) reduction of N2O by 1-hole Cu-Z is discussed and the possibility of a reaction between 2-hole Cu-Z and O-2 is considered.

Original languageEnglish
Pages (from-to)5670-5679
Number of pages10
JournalChemical science
Volume6
Issue number10
DOIs
Publication statusPublished - 2015

Keywords

  • ELECTRON-TRANSFER CENTER
  • COPPER-SULFUR CLUSTER
  • N2O REDUCTASE
  • PSEUDOMONAS-STUTZERI
  • CATALYTIC CENTER
  • SPECTROSCOPY
  • DENITRIFICATION
  • ENZYME
  • PURIFICATION
  • DINITROGEN

Cite this