- Synthesis of aromatic 13C/2H-α-ketoacid precursors to be used in selective phenylalanine and tyrosine protein labelling
-
Recent progress in protein NMR spectroscopy revealed aromatic residues to be valuable information sources for performing structure and motion analysis of high molecular weight proteins. However, the applied NMR experiments require tailored isotope labelling patterns in order to regulate spin-relaxation pathways and optimize magnetization transfer. We introduced a methodology to use α-ketoacids as metabolic amino acid precursors in cell-based overexpression of phenylalanine and/or tyrosine labelled proteins in a recent publication, which we have now developed further by providing synthetic routes to access the corresponding side-chain labelled precursors. The target compounds allow for selective introduction of 13C-1H spin systems in a highly deuterated chemical environment and feature alternating 12C-13C-12C ring-patterns. The resulting isotope distribution is especially suited to render straightforward 13C spin relaxation experiments possible, which provide insight into the dynamic properties of the corresponding labelled proteins.
- Lichtenecker, R. J.
-
-
Read Online
- Anthranilic acid, the new player in the ensemble of aromatic residue labeling precursor compounds
-
The application of metabolic precursors for selective stable isotope labeling of aromatic residues in cell-based protein overexpression has already resulted in numerous NMR probes to study the structural and dynamic characteristics of proteins. With anthranilic acid, we present the structurally simplest precursor for exclusive tryptophan side chain labeling. A synthetic route to 13C, 2H isotopologues allows the installation of isolated 13C–1H spin systems in the indole ring of tryptophan, representing a versatile tool to investigate side chain motion using relaxation-based experiments without the loss of magnetization due to strong 1JCC and weaker 2JCH scalar couplings, as well as dipolar interactions with remote hydrogens. In this article, we want to introduce this novel precursor in the context of hitherto existing techniques of in vivo aromatic residue labeling.
- Sch?rghuber, Julia,Geist, Leonhard,Bisaccia, Marilena,Weber, Frederik,Konrat, Robert,Lichtenecker, Roman J.
-
-
- Synthesis of selectively 13C-labelled benzoic acid for nuclear magnetic resonance spectroscopic measurement of glycine conjugation activity
-
The synthesis of [4-13C]benzoic acid (BA) labelled in a single protonated carbon, for use as a probe to measure glycine conjugation activity by nuclear magnetic resonance (NMR) spectroscopy, has been reported. The labelled compound was prepared
- Akira,Hasegawa,Baba
-
p. 845 - 853
(2007/10/02)
-
- Synthesis and NMR spectroscopy of stable isotope-labelled phenols and L-tyrosines
-
The syntheses of (17O)phenol from (17O)water, (18O)phenol from (18O)water, (1-13C)-phenol and (4-13C)phenol from (2-13C)acetone and (2-13C)phenol and (3-13C)phenol from (1-13C)acetone with high isotopic enrichment are described.The labelled phenols are converted into their corresponding L-tyrosines by the bacterium Erwinia herbicola.A full analysis of the 1H and 13C NMR spectra of phenol and L-tyrosine is reported.
- Winkel, C.,Aarts, M. W. M. M.,Heide, F. R. van der,Buitenhuis, E. G.,Lugtenburg, J.
-
p. 139 - 146
(2007/10/02)
-