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

    Fingerprint

    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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    AU - Bruner, Steven D.

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