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

J Biomol NMR. 2024 Sep;78(3):139-147. doi: 10.1007/s10858-024-00440-z. Epub 2024 Mar 21.

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.

MeSH terms

  • Carbon Isotopes*
  • Deuterium*
  • Escherichia coli* / metabolism
  • Isotope Labeling* / methods
  • Nuclear Magnetic Resonance, Biomolecular* / methods
  • Phenylalanine* / chemistry

Substances

  • Phenylalanine
  • Carbon Isotopes
  • Deuterium