Protein Synthesis via Activated Cysteine-Directed Protein Ligation. Academic Article

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.

authors

• Liu, Wenshe

published proceedings

• Methods Mol Biol

altmetric score

• 0.25

author list (cited authors)

• Yu, G. e., Qiao, Y., Blankenship, L. R., & Liu, W. R.

citation count

• 0

complete list of authors

• Yu, Ge||Qiao, Yuchen||Blankenship, Lauren R||Liu, Wenshe Ray

publication date

• January 2022

publisher

• Springer Nature  Publisher

published in

• Methods in molecular biology (Clifton, N.J.)  Journal

keywords

• Activated Cysteine
• C-terminus Functionalization
• Cysteine
• Esi-ms
• Expressed Protein Ligation
• Histones
• Nucleophilic Acyl Substitution
• Nucleosomes
• Protein Biosynthesis
• RNA

Digital Object Identifier (DOI)

• 10.1007/978-1-0716-2489-0_11

start page

• 159

end page

• 167

volume

• 2530

URL

• http://dx.doi.org/10.1007/978-1-0716-2489-0_11