RESUMO
In a proof-of-concept study, a mechano-chromic hydrogel was synthesized here, via chemoenzymatic click conjugation of fluorophore-labeled fibronectin into a synthetic hydrogel co-polymers (i.e., poly-N-isopropylacrylamide/polyethylene glycol). The optical FRET response could be tuned by macroscopic stretch.
Assuntos
Resinas Acrílicas/química , Fibronectinas/química , Hidrogéis/química , Polietilenoglicóis/química , Transferência Ressonante de Energia de Fluorescência , Corantes Fluorescentes/química , Fenômenos MecânicosRESUMO
To determine the binding-site of a combinatorially-selected peptide possessing a fluoroprobe, a novel cysteine reactive small photo-crosslinker that can be excited by a conventional long-wavelength ultraviolet handlamp (365 nm) was synthesized via Suzuki coupling with three steps. The crosslinker is rationally designed, not only as a bioisostere of the fluoroprobe, but as a caged-fluorophore, and the photo-crosslinked target protein became fluorescent with a large Stokes-shift. By introducing the crosslinker to a designated sulfhydryl (SH) group of a combinatorially-selected peptide, the protein-binding site of the targeted peptide was deduced by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)/fluorescence imaging followed by matrix-assisted laser desorption ionization-time of flight tandem mass spectrometry (MALDI-TOF-MS/MS) analysis.