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Review
. 2010 Dec;14(6):774-80.
doi: 10.1016/j.cbpa.2010.09.013. Epub 2010 Nov 9.

Residue-specific incorporation of non-canonical amino acids into proteins: recent developments and applications

Jeremiah A Johnson  1 , Ying Y Lu, James A Van Deventer, David A Tirrell
Affiliations
Review

Residue-specific incorporation of non-canonical amino acids into proteins: recent developments and applications

Jeremiah A Johnson et al. Curr Opin Chem Biol. 2010 Dec.

Abstract

Residue-specific incorporation of non-canonical amino acids into proteins allows facile alteration and enhancement of protein properties. In this review, we describe recent technical developments and applications of residue-specific incorporation to problems ranging from elucidation of biochemical mechanisms to engineering of protein-based biomaterials. We hope to inform the reader of the ease and broad utility of residue-specific non-canonical amino acid incorporation with the goal of inspiring investigators outside the field to consider applying this tool to their own research.

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Figures

Figure 1
Figure 1
1A: Schematic for residue-specific incorporation of non-canonical amino acids (ncAAs) into proteins. A natural mRNA contains codons for the 20 canonical amino acids (cAAs). A cAA (purple sphere) assigned to one of those codons (yellow) is replaced with an ncAA (orange star). A medium shift is performed to remove the cAA to be replaced (purple sphere) and to introduce the ncAA (orange star) along with the remaining 19 cAAs (green spheres). The ncAA is charged to the appropriate tRNA (red) by either the wild-type or a mutant aminoacyl-tRNA synthetase (aaRS). The correctly aminoacylated tRNAcAA (blue with green sphere) and the misacylated tRNA (red with orange star) are processed by the ribosome to give a globally modified protein. The left path depicts normal protein synthesis with 20 cAAs for comparison. 1B: Chemical structures for the ncAAs discussed in the text.
Figure 2
Figure 2
Schematic depictions of BONCAT and FUNCAT technologies. Both begin with metabolic labeling of newly synthesized proteins using an ncAA bearing a reactive handle (blue sphere). Then, a chemoselective reaction is performed (e.g. click chemistry) to append either an enrichment tag (BONCAT) or a fluorophore for visualization (FUNCAT). Taken together, these technologies allow simultaneous imaging and identification of new proteins in cells; proteomic responses to stimuli (changing conditions, infection, cell-cell interaction, etc.) can be monitored.
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References

    1. Munier R, Cohen GN. Incorporation of structural analogs of amino acids in bacterial proteins. Biochim. Biophys. Acta. 1956;21:592–593. - PubMed
    1. Hendrickson WA, Horton JR, LeMaster DM. Selenomethionyl proteins produced for analysis by multiwavelength anomalous diffraction (MAD): a vehicle for direct determination of three-dimensional structure. Embo J. 1990;9:1665–1672. - PMC - PubMed
    1. Yuksel D, Pamuk D, Ivanova Y, Kumar K. Protein engineering using noncanonical amino acids. Protein Eng. Des. 2010;205:221.
    1. Young TS, Schultz PG. Beyond the Canonical 20 Amino Acids: Expanding the Genetic Lexicon. J. Biol. Chem. 2010;285:11039–11044. - PMC - PubMed
    1. Voloshchuk N, Montclare JK. Incorporation of unnatural amino acids for synthetic biology. Mol. BioSyst. 2010;6:65–80. - PubMed

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