Rapid biosensor development using plant hormone receptors as reprogrammable scaffolds

Jesús Beltrán; Paul J Steiner; Matthew Bedewitz; Shuang Wei; Francis C Peterson; Zongbo Li; Brigid E Hughes; Zachary Hartley; Nicholas R Robertson; Angélica V Medina-Cucurella; Zachary T Baumer; Alison C Leonard; Sang-Youl Park; Brian F Volkman; Dmitri A Nusinow; Wenwan Zhong; Ian Wheeldon; Sean R Cutler; Timothy A Whitehead

doi:10.1038/s41587-022-01364-5

Rapid biosensor development using plant hormone receptors as reprogrammable scaffolds

Nat Biotechnol. 2022 Dec;40(12):1855-1861. doi: 10.1038/s41587-022-01364-5. Epub 2022 Jun 20.

Authors

Jesús Beltrán^#^{1

2}, Paul J Steiner^#³, Matthew Bedewitz^#³, Shuang Wei^#⁴, Francis C Peterson⁵, Zongbo Li⁶, Brigid E Hughes⁵, Zachary Hartley^{1

2}, Nicholas R Robertson⁷, Angélica V Medina-Cucurella⁸, Zachary T Baumer³, Alison C Leonard³, Sang-Youl Park¹, Brian F Volkman⁵, Dmitri A Nusinow⁹, Wenwan Zhong⁶, Ian Wheeldon^{10

11}, Sean R Cutler^{12

13

14}, Timothy A Whitehead¹⁵

Affiliations

¹ Department of Botany and Plant Sciences, University of California, Riverside, Riverside, CA, USA.
² Institute for Integrative Genome Biology, University of California, Riverside, Riverside, CA, USA.
³ Department of Chemical and Biological Engineering, University of Colorado Boulder, Boulder, CO, USA.
⁴ Department of Biochemistry, University of California, Riverside, Riverside, CA, USA.
⁵ Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI, USA.
⁶ Department of Chemistry, University of California, Riverside, Riverside, CA, USA.
⁷ Department of Bioengineering, University of California, Riverside, Riverside, USA.
⁸ Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing, MI, USA.
⁹ Donald Danforth Plant Science Center, St. Louis, MO, USA.
¹⁰ Institute for Integrative Genome Biology, University of California, Riverside, Riverside, CA, USA. wheeldon@ucr.edu.
¹¹ Department of Chemical and Environmental Engineering, University of California, Riverside, Riverside, CA, USA. wheeldon@ucr.edu.
¹² Department of Botany and Plant Sciences, University of California, Riverside, Riverside, CA, USA. cutler@ucr.edu.
¹³ Institute for Integrative Genome Biology, University of California, Riverside, Riverside, CA, USA. cutler@ucr.edu.
¹⁴ Center for Plant Cell Biology, University of California, Riverside, Riverside, CA, USA. cutler@ucr.edu.
¹⁵ Department of Chemical and Biological Engineering, University of Colorado Boulder, Boulder, CO, USA. timothy.whitehead@colorado.edu.

^# Contributed equally.

Abstract

A general method to generate biosensors for user-defined molecules could provide detection tools for a wide range of biological applications. Here, we describe an approach for the rapid engineering of biosensors using PYR1 (Pyrabactin Resistance 1), a plant abscisic acid (ABA) receptor with a malleable ligand-binding pocket and a requirement for ligand-induced heterodimerization, which facilitates the construction of sense-response functions. We applied this platform to evolve 21 sensors with nanomolar to micromolar sensitivities for a range of small molecules, including structurally diverse natural and synthetic cannabinoids and several organophosphates. X-ray crystallography analysis revealed the mechanistic basis for new ligand recognition by an evolved cannabinoid receptor. We demonstrate that PYR1-derived receptors are readily ported to various ligand-responsive outputs, including enzyme-linked immunosorbent assay (ELISA)-like assays, luminescence by protein-fragment complementation and transcriptional circuits, all with picomolar to nanomolar sensitivity. PYR1 provides a scaffold for rapidly evolving new biosensors for diverse sense-response applications.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, N.I.H., Extramural

MeSH terms

Arabidopsis Proteins* / genetics
Arabidopsis*
Biosensing Techniques*
Ligands
Plant Growth Regulators
Plants

Substances

Plant Growth Regulators
Arabidopsis Proteins
Ligands

Grants and funding

1R21DA053496-01/U.S. Department of Health & Human Services | National Institutes of Health (NIH)