Single-mutation fitness landscapes for an enzyme on multiple substrates reveal specificity is globally encoded

Emily E. Wrenbeck, Laura R. Azouz, Timothy A. Whitehead

    Research output: Contribution to journalArticle

    Abstract

    Our lack of total understanding of the intricacies of how enzymes behave has constrained our ability to robustly engineer substrate specificity. Furthermore, the mechanisms of natural evolution leading to improved or novel substrate specificities are not wholly defined. Here we generate near-comprehensive single-mutation fitness landscapes comprising >96.3% of all possible single nonsynonymous mutations for hydrolysis activity of an amidase expressed in E. coli with three different substrates. For all three selections, we find that the distribution of beneficial mutations can be described as exponential, supporting a current hypothesis for adaptive molecular evolution. Beneficial mutations in one selection have essentially no correlation with fitness for other selections and are dispersed throughout the protein sequence and structure. Our results further demonstrate the dependence of local fitness landscapes on substrate identity and provide an example of globally distributed sequence-specificity determinants for an enzyme.

    Original languageEnglish (US)
    Article number15695
    JournalNature Communications
    Volume8
    DOIs
    StatePublished - Jun 6 2017

    Profile

    mutations
    Substrate Specificity
    Mutation
    Enzymes
    Substrates
    fitness
    landscape
    enzymes
    Molecular Evolution
    Hydrolysis
    Escherichia coli
    Proteins
    amidase
    determinants
    engineers
    hydrolysis
    proteins
    Frangula
    Engineers

    ASJC Scopus subject areas

    • Chemistry(all)
    • Biochemistry, Genetics and Molecular Biology(all)
    • Physics and Astronomy(all)

    Cite this

    Single-mutation fitness landscapes for an enzyme on multiple substrates reveal specificity is globally encoded. / Wrenbeck, Emily E.; Azouz, Laura R.; Whitehead, Timothy A.

    In: Nature Communications, Vol. 8, 15695, 06.06.2017.

    Research output: Contribution to journalArticle

    Wrenbeck, Emily E.; Azouz, Laura R.; Whitehead, Timothy A. / Single-mutation fitness landscapes for an enzyme on multiple substrates reveal specificity is globally encoded.

    In: Nature Communications, Vol. 8, 15695, 06.06.2017.

    Research output: Contribution to journalArticle

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