Structure and redox properties of the diheme electron carrier cytochrome c4 from Pseudomonas aeruginosa

J Inorg Biochem. 2020 Feb;203:110889. doi: 10.1016/j.jinorgbio.2019.110889. Epub 2019 Oct 22.


At low oxygen concentrations, respiration of Pseudomonas aeruginosa (Pa) and other bacteria relies on activity of cytochrome cbb3 oxidases. A diheme cytochrome c4 (cyt c4) donates electrons to Pa cbb3 oxidases to enable oxygen reduction and proton pumping by these enzymes. Given the importance of this redox pathway for bacterial pathogenesis, both cyt c4 and cbb3 oxidase are potential targets for new antibacterial strategies. The structural information about these two proteins, however, is scarce, and functional insights for Pa and other bacteria have been primarily drawn from analyses of the analogous system from Pseudomonas stutzeri (Ps). Herein, we describe characterization of structural and redox properties of cyt c4 from Pa. The crystal structure of Pa cyt c4 has revealed that this protein is organized in two monoheme domains. The interdomain interface is more hydrophobic in Pa cyt c4, and the protein surface does not show the dipolar distribution of charges found in Ps cyt c4. The reduction potentials of the two hemes are similar in Pa cyt c4 but differ by about 100 mV in Ps cyt c4. Analyses of structural models of these and other cyt c4 proteins suggest that multiple factors contribute to the potential difference of the two hemes in these proteins, including solvent accessibility of the heme group, the distribution of surface charges, and the nature of the interdomain interface. The distinct properties of cyt c4 proteins from closely-related Pa and Ps bacteria emphasize the importance of examining the cbb3/cyt c4 redox pathway in multiple species.

Keywords: Electron transfer; Multiheme proteins; Redox partners; Reduction potentials; cbb(3) oxidase.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / metabolism
  • Catalytic Domain
  • Cytochrome c Group / chemistry*
  • Cytochrome c Group / metabolism
  • Electrons*
  • Heme / chemistry
  • Heme / metabolism
  • Iron / chemistry
  • Iron / metabolism
  • Molecular Dynamics Simulation
  • Oxidation-Reduction
  • Protein Binding
  • Pseudomonas aeruginosa / enzymology


  • Bacterial Proteins
  • Cytochrome c Group
  • cytochrome C4
  • Heme
  • Iron