Metalloprotein switches that display chemical-dependent electron transfer in cells

Nat Chem Biol. 2019 Feb;15(2):189-195. doi: 10.1038/s41589-018-0192-3. Epub 2018 Dec 17.

Abstract

Biological electron transfer is challenging to directly regulate using environmental conditions. To enable dynamic, protein-level control over energy flow in metabolic systems for synthetic biology and bioelectronics, we created ferredoxin logic gates that utilize transcriptional and post-translational inputs to control energy flow through a synthetic electron transfer pathway that is required for bacterial growth. These logic gates were created by subjecting a thermostable, plant-type ferredoxin to backbone fission and fusing the resulting fragments to a pair of proteins that self-associate, a pair of proteins whose association is stabilized by a small molecule, and to the termini of a ligand-binding domain. We show that the latter domain insertion design strategy yields an allosteric ferredoxin switch that acquires an oxygen-tolerant [2Fe-2S] cluster and can use different chemicals, including a therapeutic drug and an environmental pollutant, to control the production of a reduced metabolite in Escherichia coli and cell lysates.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Electron Spin Resonance Spectroscopy / methods
  • Electron Transport / drug effects
  • Electron Transport / physiology*
  • Electrons
  • Escherichia coli / metabolism
  • Ferredoxins / physiology
  • Metalloproteins / genetics
  • Metalloproteins / physiology*
  • Mutagenesis, Site-Directed / methods
  • Protein Processing, Post-Translational / physiology

Substances

  • Ferredoxins
  • Metalloproteins