Streamlined Synthesis and Assembly of a Hybrid Sensing Architecture with Solid Binding Proteins and Click Chemistry

• Published in: Journal of the American Chemical Society 2017-03, Vol.139 (11), p.3958-3961
• Main Authors: Swift, Brian J. F, Shadish, Jared A, DeForest, Cole A, Baneyx, François
• Format: Article
• Language: English
• Subjects: Binding Sites; Chloramphenicol - chemistry; Click Chemistry; DNA - chemistry; Green Fluorescent Proteins - chemistry; Manganese - chemistry; Models, Molecular; Particle Size; Protein Engineering; Silicon Dioxide - chemistry; Sulfides - chemistry; Surface Properties; Zinc Compounds - chemistry
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/jacs.7b00519

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 SiO2 particles 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.