title: Streamlined Synthesis and Assembly of a Hybrid Sensing Architecture with Solid Binding Proteins and Click Chemistry.
abstract: Combining bioorthogonal chemistry with the use of proteins engineered with adhesive and morphogenetic solid-binding peptides is a promising route for synthesizing hybrid materials with the economy and efficiency of living systems. Using optical sensing of chloramphenicol as a proof of concept, we show here that a GFP variant engineered with zinc sulfide and silica-binding peptides on opposite sides of its β-barrel supports the fabrication of protein-capped ZnS:Mn nanocrystals that exhibit the combined emission signatures of organic and inorganic fluorophores. Conjugation of a chloramphenicol-specific DNA aptamer to the protein shell through strain-promoted azide-alkyne cycloaddition and spontaneous concentration of the resulting nanostructures onto SiO2particles mediated by the silica-binding sequence enables visual detection of environmentally and clinically relevant concentrations of chloramphenicol through analyte-mediated inner filtering of sub-330 nm excitation light.
authors.affiliation: Department of Chemical Engineering, Box 351750, University of Washington , Seattle, Washington 98195, United States.
authors.forename: Brian J F
authors.initials: BJ
authors.lastname: Swift
authors.affiliation: Department of Chemical Engineering, Box 351750, University of Washington , Seattle, Washington 98195, United States.
authors.forename: Jared A
authors.initials: JA
authors.lastname: Shadish
authors.affiliation: Department of Chemical Engineering, Box 351750, University of Washington , Seattle, Washington 98195, United States.
authors.forename: Cole A
authors.initials: CA
authors.lastname: DeForest
authors.affiliation: Department of Chemical Engineering, Box 351750, University of Washington , Seattle, Washington 98195, United States.
authors.forename: François
authors.identifier: 0000-0001-5596-7903
authors.initials: F
authors.lastname: Baneyx
mesh_terms: D001665:Binding Sites; D002701:Chloramphenicol / Q000737:chemistry*; D057930:Click Chemistry; D004247:DNA / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000737:chemistry*; D008345:Manganese / Q000737:chemistry*; D008958:Models, Molecular; D010316:Particle Size; D015202:Protein Engineering; D012822:Silicon Dioxide / Q000737:chemistry*; D013440:Sulfides / Q000737:chemistry*; D013499:Surface Properties; D017967:Zinc Compounds / Q000737:chemistry*
chemical_list: D013440:Sulfides; D017967:Zinc Compounds; D049452:Green Fluorescent Proteins; D008345:Manganese; D002701:Chloramphenicol; D012822:Silicon Dioxide; D004247:DNA; C031238:zinc sulfide