Decorating phenylalanine side-chains with triple labeled 13C/19F/2H isotope patterns

Giorgia Toscano; Julian Holzinger; Benjamin Nagl; Georg Kontaxis; Hanspeter Kählig; Robert Konrat; Roman J. Lichtenecker

doi:10.1007/s10858-024-00440-z

Decorating phenylalanine side-chains with triple labeled ¹³C/¹⁹F/²H isotope patterns

Giorgia Toscano, Julian Holzinger, Benjamin Nagl, Georg Kontaxis, Hanspeter Kählig, Robert Konrat, Roman J. Lichtenecker (Corresponding author)

Publications: Contribution to journal › Article › Peer Reviewed

Abstract

We present an economic and straightforward method to introduce ¹³C-¹⁹F spin systems into the deuterated aromatic side chains of phenylalanine as reporters for various protein NMR applications. The method is based on the synthesis of [4-¹³C, 2,3,5,6-²H₄] 4-fluorophenylalanine from the commercially available isotope sources [2-¹³C] acetone and deuterium oxide. This compound is readily metabolized by standard Escherichia coli overexpression in a glyphosate-containing minimal medium, which results in high incorporation rates in the corresponding target proteins.

Original language	English
Journal	Journal of Biomolecular NMR
DOIs	https://doi.org/10.1007/s10858-024-00440-z
Publication status	E-pub ahead of print - Mar 2024

Austrian Fields of Science 2012

104015 Organic chemistry
106023 Molecular biology
104026 Spectroscopy

Keywords

F-TROSY
Fluorine NMR
Fluorophenylalanine
Isotope labeling
Protein overexpression

Access to Document

10.1007/s10858-024-00440-zLicence: CC BY 4.0

https://phaidra.univie.ac.at/o:2085349Licence: CC BY 4.0

Cite this

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title = "Decorating phenylalanine side-chains with triple labeled 13C/19F/2H isotope patterns",

abstract = "We present an economic and straightforward method to introduce 13C-19F spin systems into the deuterated aromatic side chains of phenylalanine as reporters for various protein NMR applications. The method is based on the synthesis of [4-13C, 2,3,5,6-2H4] 4-fluorophenylalanine from the commercially available isotope sources [2-13C] acetone and deuterium oxide. This compound is readily metabolized by standard Escherichia coli overexpression in a glyphosate-containing minimal medium, which results in high incorporation rates in the corresponding target proteins.",

keywords = "F-TROSY, Fluorine NMR, Fluorophenylalanine, Isotope labeling, Protein overexpression",

author = "Giorgia Toscano and Julian Holzinger and Benjamin Nagl and Georg Kontaxis and Hanspeter K{\"a}hlig and Robert Konrat and Lichtenecker, {Roman J.}",

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T1 - Decorating phenylalanine side-chains with triple labeled 13C/19F/2H isotope patterns

AU - Toscano, Giorgia

AU - Holzinger, Julian

AU - Nagl, Benjamin

AU - Kontaxis, Georg

AU - Kählig, Hanspeter

AU - Konrat, Robert

AU - Lichtenecker, Roman J.

N1 - Accession Number WOS:001190138000001 PubMed ID 38509441

PY - 2024/3

Y1 - 2024/3

N2 - We present an economic and straightforward method to introduce 13C-19F spin systems into the deuterated aromatic side chains of phenylalanine as reporters for various protein NMR applications. The method is based on the synthesis of [4-13C, 2,3,5,6-2H4] 4-fluorophenylalanine from the commercially available isotope sources [2-13C] acetone and deuterium oxide. This compound is readily metabolized by standard Escherichia coli overexpression in a glyphosate-containing minimal medium, which results in high incorporation rates in the corresponding target proteins.

AB - We present an economic and straightforward method to introduce 13C-19F spin systems into the deuterated aromatic side chains of phenylalanine as reporters for various protein NMR applications. The method is based on the synthesis of [4-13C, 2,3,5,6-2H4] 4-fluorophenylalanine from the commercially available isotope sources [2-13C] acetone and deuterium oxide. This compound is readily metabolized by standard Escherichia coli overexpression in a glyphosate-containing minimal medium, which results in high incorporation rates in the corresponding target proteins.

KW - F-TROSY

KW - Fluorine NMR

KW - Fluorophenylalanine

KW - Isotope labeling

KW - Protein overexpression

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