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- W4328108973 abstract "<strong class=journal-contentHeaderColor>Abstract.</strong> The novel eCell system maintains the activity of the entire repertoire of metabolic <em>E. coli</em> enzymes in cell-free protein synthesis. We show that this can be harnessed to produce proteins with selectively <sup>13</sup>C-labelled amino acids from inexpensive <sup>13</sup>C-labelled precursors. The system is demonstrated with selective <sup>13</sup>C-labelling of methyl groups in the proteins ubiquitin and peptidyl-prolyl <em>cis–trans</em> isomerase B. Starting from 3-<sup>13</sup>C-pyruvate, <sup>13</sup>C-HSQC cross-peaks are obtained devoid of one-bond <sup>13</sup>C-<sup>13</sup>C scalar couplings. Starting from 2-<sup>13</sup>C-methyl-acetolactate, single methyl groups of valine and leucine are labelled. Labelling efficiencies are 70 % or higher, and the method allows to produce perdeuterated proteins with protonated methyl groups in residue-selective manner. The system uses the isotope-labelled precursors sparingly and is more readily scalable than conventional cell-free systems." @default.
- W4328108973 created "2023-03-22" @default.
- W4328108973 creator A5004707292 @default.
- W4328108973 date "2023-03-21" @default.
- W4328108973 modified "2023-09-25" @default.
- W4328108973 title "Comment on mr-2023-3" @default.
- W4328108973 doi "https://doi.org/10.5194/mr-2023-3-rc1" @default.
- W4328108973 hasPublicationYear "2023" @default.
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