Enzyme-catalyzed expressed protein ligation

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

doi:10.1038/nmeth.4004

Enzyme-catalyzed expressed protein ligation

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

School of Medicine

Research output: Contribution to journal › Article › peer-review

30 Scopus citations

Abstract

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 language	English (US)
Pages (from-to)	925-927
Number of pages	3
Journal	Nature Methods
Volume	13
Issue number	11
DOIs	https://doi.org/10.1038/nmeth.4004
State	Published - Nov 1 2016

ASJC Scopus subject areas

Biotechnology
Biochemistry
Molecular Biology
Cell Biology

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AU - Henager, Samuel H.

AU - Chu, Nam

AU - Chen, Zan

AU - Bolduc, David

AU - Dempsey, Daniel R.

AU - Hwang, Yousang

AU - Wells, James

AU - Cole, Philip A.

PY - 2016/11/1

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AB - 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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Enzyme-catalyzed expressed protein ligation

Abstract

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