AI Article Synopsis
- Fluorosubstituted tryptophans, like 7-fluoro-tryptophan, are useful in studying proteins through fluorescence and NMR techniques.
- In cyclophilin A, UV exposure of 7-fluoro-tryptophan leads to the creation of a bright fluorophore by crosslinking with nearby phenylalanine, achieving a high quantum yield and fluorescence lifetime.
- This study is the first to document a Phe-Trp crosslink in a protein, potentially paving the way for new fluorescence probes and methods to utilize aromatic crosslinks in protein research.
Article Abstract
Fluorosubstituted tryptophans serve as valuable probes for fluorescence and nuclear magnetic resonance (NMR) studies of proteins. Here, we describe an unusual photoreactivity introduced by replacing the single tryptophan in cyclophilin A with 7-fluoro-tryptophan. UV exposure at 282 nm defluorinates 7-fluoro-tryptophan and crosslinks it to a nearby phenylalanine, generating a bright fluorophore. The crosslink-containing fluorescent protein possesses a large quantum yield of ∼0.40 with a fluorescence lifetime of 2.38 ns. The chemical nature of the crosslink and the three-dimensional protein structure were determined by mass spectrometry and NMR spectroscopy. To the best of our knowledge, this is the first report of a Phe-Trp crosslink in a protein. Our finding may break new ground for developing novel fluorescence probes and for devising new strategies to exploit aromatic crosslinks in proteins.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9233106 | PMC |
| http://dx.doi.org/10.1021/jacs.2c02054 | DOI Listing |
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