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Rapid biosensor development using plant hormone receptors as reprogrammable scaffolds.
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.
Afiliação
  • Beltrán J; Department of Botany and Plant Sciences, University of California, Riverside, Riverside, CA, USA.
  • Steiner PJ; Institute for Integrative Genome Biology, University of California, Riverside, Riverside, CA, USA.
  • Bedewitz M; Department of Chemical and Biological Engineering, University of Colorado Boulder, Boulder, CO, USA.
  • Wei S; Department of Chemical and Biological Engineering, University of Colorado Boulder, Boulder, CO, USA.
  • Peterson FC; Department of Biochemistry, University of California, Riverside, Riverside, CA, USA.
  • Li Z; Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI, USA.
  • Hughes BE; Department of Chemistry, University of California, Riverside, Riverside, CA, USA.
  • Hartley Z; Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI, USA.
  • Robertson NR; Department of Botany and Plant Sciences, University of California, Riverside, Riverside, CA, USA.
  • Medina-Cucurella AV; Institute for Integrative Genome Biology, University of California, Riverside, Riverside, CA, USA.
  • Baumer ZT; Department of Bioengineering, University of California, Riverside, Riverside, USA.
  • Leonard AC; Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing, MI, USA.
  • Park SY; Department of Chemical and Biological Engineering, University of Colorado Boulder, Boulder, CO, USA.
  • Volkman BF; Department of Chemical and Biological Engineering, University of Colorado Boulder, Boulder, CO, USA.
  • Nusinow DA; Department of Botany and Plant Sciences, University of California, Riverside, Riverside, CA, USA.
  • Zhong W; Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI, USA.
  • Wheeldon I; Donald Danforth Plant Science Center, St. Louis, MO, USA.
  • Cutler SR; Department of Chemistry, University of California, Riverside, Riverside, CA, USA.
  • Whitehead TA; Institute for Integrative Genome Biology, University of California, Riverside, Riverside, CA, USA. wheeldon@ucr.edu.
Nat Biotechnol ; 40(12): 1855-1861, 2022 12.
Article em En | MEDLINE | ID: mdl-35726092
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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Técnicas Biossensoriais / Arabidopsis / Proteínas de Arabidopsis Idioma: En Revista: Nat Biotechnol Ano de publicação: 2022 Tipo de documento: Article
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Técnicas Biossensoriais / Arabidopsis / Proteínas de Arabidopsis Idioma: En Revista: Nat Biotechnol Ano de publicação: 2022 Tipo de documento: Article