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Enzyme-catalyzed expressed protein ligation.

  • Author(s): Henager, Samuel H
  • Chu, Nam
  • Chen, Zan
  • Bolduc, David
  • Dempsey, Daniel R
  • Hwang, Yousang
  • Wells, James
  • Cole, Philip A
  • et al.

Expressed protein ligation is a valuable method for protein semisynthesis that involves the reaction of recombinant protein C-terminal thioesters with N-terminal cysteine (N-Cys)-containing peptides, but the requirement of a Cys residue at the ligation junction can limit the utility of this method. Here we employ subtiligase variants to efficiently ligate Cys-free peptides to protein thioesters. Using this method, we have more accurately determined the effect of C-terminal phosphorylation on the tumor suppressor protein PTEN.

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