Ugwumba, Isaac N; Ozawa, Kiyoshi; de la Cruz, Laura; Xu, Zhi-Qiang; Herlt, Anthony; Hadler, Kieran S; Coppin, Christopher Wayne; Brown, S; Schenk, Gerhard; Oakeshott, John Graham; Otting, Gottfried
Development of enzyme inhibitors requires an activity assay for the identification of hits and lead compounds. To determine dissociation constants in a straightforward manner, we explored the use of a genetically encoded fluorescent amino acid for site-specific tagging of the target protein. The unnatural amino acid 7-(hydroxy-coumarin-4-yl) ethylglycine (Hco) was site-specifically incorporated in the target protein by cell-free protein synthesis using an orthogonal amber suppressor...[Show more]
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