Rapid biosensor development using plant hormone receptors as reprogrammable scaffolds
Abstrak
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.
Topik & Kata Kunci
Penulis (19)
J. Beltrán
Paul J. Steiner
Matthew A. Bedewitz
Shuang Wei
F. Peterson
Zongbo Li
Brigid E Hughes
Zachary Hartley
Nicholas R. Robertson
Angélica V. Medina-Cucurella
Z. Baumer
Alison C. Leonard
Sang-Youl Park
B. Volkman
Dmitri A. Nusinow
Wenwan Zhong
I. Wheeldon
S. Cutler
Timothy A. Whitehead
Akses Cepat
- Tahun Terbit
- 2022
- Bahasa
- en
- Total Sitasi
- 67×
- Sumber Database
- Semantic Scholar
- DOI
- 10.1038/s41587-022-01364-5
- Akses
- Open Access ✓