Highly Selective Stable Isotope Labeling of Histidine Residues by Using a Novel Precursor in E. coli‐Based Overexpression Systems. (12th June 2017)

• Record Type: Journal Article
• Authors: Schörghuber, Julia; Geist, Leonhard; Platzer, Gerald; Konrat, Robert; Lichtenecker, Roman J.
• Is Part Of: Chembiochem. Volume 18:Number 15(2017)
• Page Start: 1487
• Page End: 1491
• Publication Date: 2017-06-12
• Subjects: histidine -- imidazole -- isotope labeling -- NMR spectroscopy -- protein expression; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.201700192
• Languages: English
• ISSNs: 1439-4227

Abstract

Abstract: The importance of NMR spectroscopy in unraveling the structural and dynamic properties of proteins is ever‐expanding owing to progress in experimental techniques, hardware development, and novel labeling approaches. Multiple sophisticated methods of aliphatic residue labeling can be found in the literature, whereas the selective incorporation of NMR active isotopes into other amino acids still holds the potential for improvement. In order to close this methodological gap, we present a novel metabolic precursor for cell‐based protein overexpression to assemble 13 C/ 2 H isotope patterns in the peptide backbone, as well as in side chain positions of a mechanistically distinguished histidine residue. Abstract : For histidines only ! The enol tautomer of imidazole‐4‐pyruvic acid is introduced as a novel precursor in E. coli ‐based labeling of histidine residues. A straightforward synthetic route allows for selective 13 C‐labeling at the ϵ 1 imidazole side‐chain position, thus providing a valuable tool to study histidine's dynamic properties, p K a values, and tautomer populations by protein NMR spectroscopy.