Rapid biosensor development using plant hormone receptors as reprogrammable scaffolds
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.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Jun 20, 2022
- Source ID
- 10.1038/s41587-022-01364-5
Entities
People
- Alison C. Leonard
- Angélica V. Medina-cucurella
- Brian F Volkman
- Brigid E. Hughes
- Dmitri A. Nusinow
- Francis C. Peterson
- Ian R Wheeldon
- Jesús Beltrán
- Matthew A Bedewitz
- Nicholas R. Robertson
- Paul J Steiner
- Sang-Youl Park
- Sean Cutler
- Shuang Wei
- Timothy A Whitehead
- Wenwan Zhong
- Zachary Hartley
- Zachary T. Baumer
- Zongbo Li
Organizations
- Cidara Therapeutics
- National Institutes of Health
- National Science Foundation
- United States Department of Defense