Abstract
Proteins with a functionalized C-terminus are critical to synthesizing large proteins via expressed protein ligation. To overcome the limitations of currently available C-terminus functionalization strategies, we established an approach based on a small molecule cyanylating reagent that chemically activates a cysteine in a recombinant protein at its N-side amide for undergoing nucleophilic acyl substitution with amines. We demonstrated the versatility of this approach by successfully synthesizing RNAse H with its RNA hydrolyzing activity restored and in vitro nucleosome build with a C-terminal posttranslational modified histone H2A. This technique will expand the landscape of protein chemical synthesis and its application in new research fields significantly.
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Yu, G., Qiao, Y., Blankenship, L.R., Liu, W.R. (2022). Protein Synthesis via Activated Cysteine-Directed Protein Ligation. In: Li, X. (eds) Chemical Protein Synthesis. Methods in Molecular Biology, vol 2530. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2489-0_11
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DOI: https://doi.org/10.1007/978-1-0716-2489-0_11
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