Details

Autor(en) / Beteiligte

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

Titel

Rapid biosensor development using plant hormone receptors as reprogrammable scaffolds

Ist Teil von

• Nature biotechnology, 2022-12, Vol.40 (12), p.1855-1861

Ort / Verlag

United States

Erscheinungsjahr

2022

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.