AI Article Synopsis
- A novel small photo-crosslinker was created to study the binding site of a specific peptide that contains a fluoroprobe, using a process that involves Suzuki coupling.
- The crosslinker is designed to act as a bioisostere of the fluoroprobe and can be activated under a standard UV light, turning the target protein fluorescent with a significant Stokes-shift.
- The binding site of the peptide was identified using techniques like SDS-PAGE for fluorescence imaging and MALDI-TOF-MS/MS for further analysis.
Article Abstract
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.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6274937 | PMC |
| http://dx.doi.org/10.3390/ijms19113682 | DOI Listing |
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