Specific Modulation of Protein Activity by Using a Bioorthogonal Reaction

• DOI: 10.1002/cbic.201402423
• Publication Date: 2014-09-24T20:11:45Z
• Authors: John B. Warner; Anand K. Muthusamy; E. James Petersson
• Keywords: Bioorthogonal Chemistry; Protein Engineering; Target protein

Unnatural amino acids with bioorthogonal reactive groups have the potential to provide a rapid and specific mechanism for covalently inhibiting protein of interest. Here, we use mutagenesis insert an unnatural acid containing azide group (Z) into target at positions such that "click" reaction alkyne modulator (X) will alter function protein. This bioorthogonally pair can engender specificity X Z-containing protein, even if is otherwise identical another allowing validation in living cells. We demonstrate our method using inhibition Escherichia coli enzyme aminoacyl transferase by both active-site occlusion allosteric mechanisms. termed this "clickable magic bullet" strategy, it should be generally applicable studying effects inhibition, within limits mutagenesis.