Determination of the molecular basis for coprogen import by Gram-negative bacteria

Research output: Contribution to journalArticleResearchpeer-review

Abstract

In order to survive in mixed microbial communities, some species of fungi secrete coprogens, siderophores that facilitate capture of the scarce nutrient iron. The TonB-dependent transporter FhuE is integrated in the outer membrane of Gram-negative bacteria and has been reported to scavenge these fungally produced coprogens. In this work, an Escherichia coli strain was engineered that is dependent solely on FhuE for its access to siderophore-sequestered iron. Using this tool, it is shown that while FhuE is highly active in the import of coprogens, it has some level of promiscuity, acting as a low-affinity transporter for related siderophores. The crystal structure of FhuE in complex with coprogen was determined, providing a structural basis to explain this selective promiscuity. The structural data, in combination with functional analysis, presented in this work show that FhuE has evolved to specifically engage with planar siderophores. A potential evolutionary driver, and a critical consequence of this selectivity, is that it allows FhuE to exclude antibiotics that mimic nonplanar hydroxamate siderophores: these toxic molecules could otherwise cross the outer membrane barrier through a Trojan horse mechanism.

Original languageEnglish
Pages (from-to)401-411
Number of pages11
JournalIUCrJ
Volume6
Issue number3
DOIs
Publication statusPublished - May 2019

Keywords

  • coprogen
  • Gram-negative bacteria
  • iron piracy
  • membrane proteins
  • membrane transport
  • protein structure
  • siderophores
  • structure determination
  • transporters
  • X-ray crystallography

Cite this

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title = "Determination of the molecular basis for coprogen import by Gram-negative bacteria",
abstract = "In order to survive in mixed microbial communities, some species of fungi secrete coprogens, siderophores that facilitate capture of the scarce nutrient iron. The TonB-dependent transporter FhuE is integrated in the outer membrane of Gram-negative bacteria and has been reported to scavenge these fungally produced coprogens. In this work, an Escherichia coli strain was engineered that is dependent solely on FhuE for its access to siderophore-sequestered iron. Using this tool, it is shown that while FhuE is highly active in the import of coprogens, it has some level of promiscuity, acting as a low-affinity transporter for related siderophores. The crystal structure of FhuE in complex with coprogen was determined, providing a structural basis to explain this selective promiscuity. The structural data, in combination with functional analysis, presented in this work show that FhuE has evolved to specifically engage with planar siderophores. A potential evolutionary driver, and a critical consequence of this selectivity, is that it allows FhuE to exclude antibiotics that mimic nonplanar hydroxamate siderophores: these toxic molecules could otherwise cross the outer membrane barrier through a Trojan horse mechanism.",
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author = "Rhys Grinter and Trevor Lithgow",
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language = "English",
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}

Determination of the molecular basis for coprogen import by Gram-negative bacteria. / Grinter, Rhys; Lithgow, Trevor.

In: IUCrJ, Vol. 6, No. 3, 05.2019, p. 401-411.

Research output: Contribution to journalArticleResearchpeer-review

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