Guiding Biomolecular Interactions in Cells Using de Novo Protein - Protein Interfaces

Abigail Smith, Franziska Thomas, Deborah Shoemark, Dek Woolfson*, Nigel Savery

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

14 Citations (Scopus)
64 Downloads (Pure)


An improved ability to direct and control biomolecular interactions in living cells would have an impact on synthetic biology. A key issue is the need to introduce interacting components that act orthogonally to endogenous proteomes and interactomes. Here, we show that low-complexity, de novo designed protein–protein interaction (PPI) domains can substitute for natural PPIs and guide engineered protein–DNA interactions in Escherichia coli. Specifically, we use de novo homo- and heterodimeric coiled coils to reconstitute a cytoplasmic split adenylate cyclase, recruit RNA polymerase to a promoter and activate gene expression, and oligomerize both natural and designed DNA-binding domains to repress transcription. Moreover, the stabilities of the heterodimeric coiled coils can be modulated by rational design and, thus, adjust the levels of gene activation and repression in vivo. These experiments demonstrate the possibilities for using designed proteins and interactions to control biomolecular systems such as enzyme cascades and circuits in cells.
Original languageEnglish
Pages (from-to)1284-1293
Number of pages10
JournalACS Synthetic Biology
Issue number6
Early online date6 May 2019
Publication statusPublished - 21 Jun 2019

Structured keywords

  • BrisSynBio
  • Bristol BioDesign Institute


  • transcriptional control
  • TAL effectors
  • protein−protein interaction
  • DNA−protein interaction
  • de novo protein design
  • α-helical coiled coil


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