A novel mechanism of inhibition by phenylthiourea on PvdP, a tyrosinase synthesizing pyoverdine of Pseudomonas aeruginosa

Joko P Wibowo, Fernando A Batista, Niels van Oosterwijk, Matthew R Groves, Frank J Dekker, Wim J Quax*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

4 Citations (Scopus)
203 Downloads (Pure)

Abstract

The biosynthesis of pyoverdine, the major siderophore of Pseudomonas aeruginosa, is a well-organized process involving a discrete number of enzyme-catalyzed steps. The final step of this process involves the PvdP tyrosinase, which converts ferribactin into pyoverdine. Thus, inhibition of the PvdP tyrosinase activity provides an attractive strategy to interfere with siderophore synthesis to manage P. aeruginosa infections. Here, we report phenylthiourea as a non-competitive inhibitor of PvdP for which we solved a crystal structure in complex with PvdP. The crystal structure indicates that phenylthiourea binds to an allosteric binding site and thereby interferes with its tyrosinase activity. We further provide proofs that PvdP tyrosinase inhibition by phenylthiourea requires the C-terminal lid region. This provides opportunities to develop inhibitors that target the allosteric site, which seems to be confined to fluorescent pseudomonads, and not the tyrosinase active site. Furthermore, increases the chances to identify PvdP inhibitors that selectively interfere with siderophore synthesis.

Original languageEnglish
Pages (from-to)212-221
Number of pages10
JournalInternational Journal of Biological Macromolecules
Volume146
Early online date30-Dec-2019
DOIs
Publication statusPublished - 1-Mar-2020

Keywords

  • Phenylthiourea
  • Pseudomonas aeruginosa
  • PvdP
  • Pyoverdine
  • Tyrosinase
  • aurantiin
  • kojic acid
  • mimosine
  • monophenol monooxygenase
  • phenylthiourea
  • piceatannol
  • PvdP enzyme
  • pyoverdine
  • quercetin
  • siderophore
  • stigmasterol
  • tropolone
  • unclassified drug
  • allosteric site
  • article
  • Bacillus megaterium
  • binding site
  • carboxy terminal sequence
  • centrifugation
  • controlled study
  • crystal structure
  • crystallization
  • cytokine production
  • cytoplasm
  • drug protein binding
  • enzyme active site
  • enzyme activity
  • enzyme inhibition
  • enzyme kinetics
  • IC50
  • Michaelis Menten kinetics
  • molecular cloning
  • nonhuman
  • periplasm
  • protein stability
  • structural homology
  • upregulation
  • X ray diffraction

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