Enzyme-catalyzed expressed protein ligation

Samuel H. Henager, Nam Chu, Zan Chen, David Bolduc, Daniel R. Dempsey, Yousang Hwang, James Wells, Philip A. Cole

Research output: Contribution to journalArticlepeer-review

30 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)925-927
Number of pages3
JournalNature Methods
Issue number11
StatePublished - Nov 1 2016

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology


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