Structure and Functional Analysis of ClbQ, an Unusual Intermediate-Releasing Thioesterase from the Colibactin Biosynthetic Pathway

Naga Sandhya Guntaka, Alan Healy, Jason M. Crawford, Seth B. Herzon, Steven D. Bruner

Research output: Contribution to journalArticle

Abstract

Colibactin is a genotoxic hybrid nonribosomal peptide/polyketide secondary metabolite produced by various pathogenic and probiotic bacteria residing in the human gut. The presence of colibactin metabolites has been correlated to colorectal cancer formation in several studies. The specific function of many gene products in the colibactin gene cluster can be predicted. However, the role of ClbQ, a type II editing thioesterase, has not been established. The importance of ClbQ has been demonstrated by genetic deletions that abolish colibactin cytotoxic activity, and recent studies suggest an atypical role in releasing pathway intermediates from the assembly line. Here we report the 2.0 Å crystal structure and biochemical characterization of ClbQ. Our data reveal that ClbQ exhibits greater catalytic efficiency toward acyl-thioester substrates as compared to precolibactin intermediates and does not discriminate among carrier proteins. Cyclized pyridone-containing colibactins, which are off-pathway derivatives, are not viable substrates for ClbQ, while linear precursors are, supporting a role of ClbQ in facilitating the promiscuous off-loading of premature precolibactin metabolites and novel insights into colibactin biosynthesis.

Original languageEnglish (US)
Pages (from-to)2598-2608
Number of pages11
JournalACS Chemical Biology
Volume12
Issue number10
DOIs
StatePublished - Oct 20 2017

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Functional analysis
Biosynthetic Pathways
Metabolites
Genes
Pyridones
Polyketides
Biosynthesis
Probiotics
Substrates
Multigene Family
colibactin
Colorectal Neoplasms
Bacteria
Carrier Proteins
Crystal structure
Derivatives
Peptides

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

Cite this

Structure and Functional Analysis of ClbQ, an Unusual Intermediate-Releasing Thioesterase from the Colibactin Biosynthetic Pathway. / Guntaka, Naga Sandhya; Healy, Alan; Crawford, Jason M.; Herzon, Seth B.; Bruner, Steven D.

In: ACS Chemical Biology, Vol. 12, No. 10, 20.10.2017, p. 2598-2608.

Research output: Contribution to journalArticle

Guntaka, Naga Sandhya ; Healy, Alan ; Crawford, Jason M. ; Herzon, Seth B. ; Bruner, Steven D. / Structure and Functional Analysis of ClbQ, an Unusual Intermediate-Releasing Thioesterase from the Colibactin Biosynthetic Pathway. In: ACS Chemical Biology. 2017 ; Vol. 12, No. 10. pp. 2598-2608.
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