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

J Am Chem Soc. 2017 Mar 22;139(11):3958-3961. doi: 10.1021/jacs.7b00519. Epub 2017 Mar 13.

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 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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • 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*

Substances

  • Sulfides
  • Zinc Compounds
  • Green Fluorescent Proteins
  • Manganese
  • Chloramphenicol
  • Silicon Dioxide
  • DNA
  • zinc sulfide