Unusually broad-spectrum small-molecule sensing using a single protein scaffold

Proc Natl Acad Sci U S A. 2025 Dec 23;122(51):e2519924122. doi: 10.1073/pnas.2519924122. Epub 2025 Dec 15.

Abstract

Small-molecule sensing in plants is dominated by chemical-induced dimerization modules. In the abscisic acid (ABA) system, allosteric receptors recruit phosphatase effectors and achieve nM in vivo responses from µM receptor-ligand interactions. This sensitivity amplification could enable ABA receptors to serve as generic scaffolds for designing small-molecule sensors. To test this, we screened collections of mutant ABA-receptors against 2,726 drugs and other ligands and identified 553 sensors for 6.6% of these ligands. The mutational patterns indicate strong selection for ligand-specific binding pockets. We used these data to develop a sensor design pipeline and isolated sensors for multiple plant natural products, 2,4,6-trinitrotoluene (TNT), and "forever" per- and polyfluoroalkyl substances (PFAS). Thus, the ABA sensor system enables design and isolation of small-molecule sensors with broad chemical scope and antibody-like simplicity.

Keywords: PFAS; biosensors; chemical induced dimerization; plant hormone receptors; synthetic biology.

MeSH terms

  • Abscisic Acid / metabolism
  • Arabidopsis Proteins* / chemistry
  • Arabidopsis Proteins* / genetics
  • Arabidopsis Proteins* / metabolism
  • Arabidopsis* / genetics
  • Arabidopsis* / metabolism
  • Biosensing Techniques* / methods
  • Ligands
  • Small Molecule Libraries* / chemistry
  • Trinitrotoluene

Substances

  • Ligands
  • Abscisic Acid
  • Trinitrotoluene
  • Small Molecule Libraries
  • Arabidopsis Proteins