Synthesis and In Vivo Incorporation of 4-Difluoromethyl-L-Phenylalanine Into Proteins.

Authors

Daniel S Honeycutt, Jason M Mrosla, Sarah A Sexton, Giovanni E Cavalli, Emmerson G Bartels, Brian Pallares, William K McCarthy, Ruojun Wu, Fang Wang, Jacob M Goldberg

Published in

Chembiochem : a European journal of chemical biology. Volume 27. Issue 18. Pages e70541. Sep 28, 2026.

Abstract

The difluoromethyl group exhibits unique chemical and nuclear magnetic resonance (NMR) spectroscopic properties. Here, the synthesis and characterization of 4-difluoromethyl-L-phenylalanine is reported. Methods have been developed to incorporate this versatile unnatural amino acid into full-length green fluorescent protein and carbonic anhydrase, using an orthogonal transfer RNA (tRNA)/synthetase pair with standard genetic code expansion techniques. Taking advantage of the characteristic ¹⁹F NMR behavior of the difluoromethyl group, the potential application of the difluoromethyl group as a probe in protein NMR studies was explored.

PMID:
42775918
Bibliographic data and abstract were imported from PubMed on 23 Sep 2026.

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