Introducing Bioorthogonal Functionalities into Proteins in Living Cells

• DOI: 10.1021/ar200067r
• Publication Date: 2011-06-03T10:29:16Z
• Authors: Ziyang Hao; Senlian Hong; Xing Chen; Peng R. Chen
• Keywords: Bioorthogonal Chemistry

Proteins are the workhorses of cell, playing crucial roles in virtually every biological process. The revolutionary ability to visualize and monitor proteins living systems, which is largely result development green fluorescence protein (GFP) its derivatives, has dramatically expanded our understanding dynamics function. Still, GFPs ill suited many circumstances; one major drawback their relatively large size, can significantly perturb functions native they fused.To bridge this gap, scientists working at chemistry–biology interface have developed methods install bioorthogonal functional groups into cells. group is, by definition, a non-native nonperturbing chemical group. But more importantly, installed handle able react with probe bearing complementary functionality highly selective fashion cell operating physiological state. Although extensive efforts been directed toward reactions, introducing functionalities systems remains an ongoing challenge. In Account, we survey recent progress area, focusing on genetic code expansion approach.In nature, uses posttranslational modifications append necessary that beyond those contained canonical 20 amino acids. Taking lessons from chosen or engineered certain enzymes modify target handles. Alternatively, use cell's translational machinery genetically encode functionalities, typically form unnatural acids (UAAs), proteins; be done residue-specific site-specific manner. For studying function cells, modification means usually favored.A variety UAAs as well other encoded interest. approach established bacteria, tagging mammalian cells challenging. A facile pyrrolysine-based system, might potentially become "one-stop shop" for both prokaryotic eukaryotic recently emerged. This technology effectively introduce series handles subsequent conjugation small-molecule probes. Moreover, method may provide precise labeling than GFP tagging. These advancements build foundation complex cellular processes, such important receptors surfaces.