Structural and functional characterization of itaconyl-CoA hydratase and citramalyl-CoA lyase involved in itaconate metabolism of Pseudomonas aeruginosa

Structure. 2024 Jul 11;32(7):941-952.e3. doi: 10.1016/j.str.2024.04.004. Epub 2024 Apr 26.

Abstract

Itaconate is a key anti-inflammatory/antibacterial metabolite in pathogen-macrophage interactions that induces adaptive changes in Pseudomonas aeruginosa-exposed airways. However, the impact and mechanisms underlying itaconate metabolism remain unclear. Our study reveals that itaconate significantly upregulates the expression of pyoverdine in P. aeruginosa and enhances its tolerance to tobramycin. Notably, the enzymes responsible for efficient itaconate metabolism, PaIch and PaCcl, play crucial roles in both utilizing itaconate and clearing its toxic metabolic intermediates. By using protein crystallography and molecular dynamics simulations analyses, we have elucidated the unique catalytic center and substrate-binding pocket of PaIch, which contribute to its highly efficient catalysis. Meanwhile, analysis of PaCcl has revealed how interactions between domains regulate the conformational changes of the active sites and binding pockets, influencing the catalytic process. Overall, our research uncovers the significance and mechanisms of PaIch and PaCcl in the efficient metabolism of itaconate by P. aeruginosa.

Keywords: PaCcl; PaIch; Pseudomonas aeruginosa; itaconate metabolism; molecular dynamics.

MeSH terms

  • Bacterial Proteins* / chemistry
  • Bacterial Proteins* / genetics
  • Bacterial Proteins* / metabolism
  • Binding Sites
  • Catalytic Domain*
  • Crystallography, X-Ray
  • Hydro-Lyases / chemistry
  • Hydro-Lyases / genetics
  • Hydro-Lyases / metabolism
  • Molecular Dynamics Simulation
  • Oxo-Acid-Lyases* / chemistry
  • Protein Binding
  • Pseudomonas aeruginosa* / enzymology
  • Pseudomonas aeruginosa* / metabolism
  • Substrate Specificity
  • Succinates* / chemistry
  • Succinates* / metabolism

Substances

  • Bacterial Proteins
  • Hydro-Lyases
  • itaconic acid
  • Succinates
  • citramalate lyase
  • Oxo-Acid-Lyases