The synthesis of specifically isotope labelled fluorotryptophan and its use in mammalian cell-based protein expression for 19F-NMR applications

19F nuclei serve as versatile sensors for detecting protein interactions and dynamics in biomolecular NMR spectroscopy. Although various methods have been developed to incorporate fluorine-containing aromatic residues into proteins using E. coli or cell-free expression techniques, similar approaches...

Full description

Saved in:
Bibliographic Details
Published in:Chemical communications (Cambridge, England) England), 2024-11, Vol.60 (96), p.14188-14191
Main Authors: Toscano, Giorgia, Rosati, Martina, Barbieri, Letizia, Maier, Katharina, Banci, Lucia, Luchinat, Enrico, Konrat, Robert, Lichtenecker, Roman J
Format: Article
Language:English
Subjects:
Carbon 13
Carbonic anhydrase
Deuteration
E coli
Fluorine
Mammals
NMR spectroscopy
Proteins
Superoxide dismutase
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:19F nuclei serve as versatile sensors for detecting protein interactions and dynamics in biomolecular NMR spectroscopy. Although various methods have been developed to incorporate fluorine-containing aromatic residues into proteins using E. coli or cell-free expression techniques, similar approaches for protein production in mammalian cell lines remain limited. Here, we present a cost-effective synthetic route to obtain selectively deuterated, carbon-13 labeled fluorotryptophan and demonstrate its use in introducing 19F–13C spin pairs into carbonic anhydrase 2 and superoxide dismutase, following an expression protocol utilizing HEK cells.
ISSN:1359-7345
1364-548X
1364-548X
DOI:10.1039/d4cc04789c