Ozawa, Kiyoshi; Headlam, Madeleine; Mouradov, Dmitri; Watt, Stephen J; Beck, Jennifer; Rodgers, Kenneth; Dean, Roger; Dixon, Nicholas; Otting, Gottfried; Huber, Thomas
An Escherichia coli cell-free transcription/translation system was used to explore the high-level incorporation of L-3,4-dihydroxyphenylalanine (DOPA) into proteins by replacing tyrosine with DOPA in the reaction mixtures. ESI-MS showed specific incorporation of DOPA in place of tyrosine. More than 90% DOPA incorporation at each tyrosine site was achieved, allowing the recording of clean15N-HSQC NMR spectra. A redox-staining method specific for DOPA was shown to provide a sensitive and...[Show more]
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