Improved spectral resolution of [ < sup > 13 < /sup > C, < sup > 1 < /sup > H]-HSQC spectra of aromatic amino acid residues in proteins produced by cell-free synthesis from inexpensive < sup > 13 < /sup > C-labelled precursors

J Biomol NMR. 2023 Jun 20. doi: 10.1007/s10858-023-00420-9. Online ahead of print.ABSTRACTCell-free protein synthesis using eCells allows production of amino acids from inexpensive 13C-labelled precursors. We show that the metabolic pathway converting pyruvate, glucose and erythrose into aromatic amino acids is maintained in eCells. Judicious choice of 13C-labelled starting material leads to proteins, where the sidechains of aromatic amino acids display [13C,1H]-HSQC cross-peaks free of one-bond 13C-13C couplings. Selective 13C-labelling of tyrosine and phenylalanine residues is achieved simply by using different compositions of the reaction buffers.PMID:37338652 | DOI:10.1007/s10858-023-00420-9

Source: Journal of Bimolecular NMR - June 20, 2023 Category: Biomedical Science Authors: Damian Van Raad Thomas Huber Gottfried Otting Source Type: research