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

  • 6 Citations

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.

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

Profile

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

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

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