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Nat Methods. 2016 Nov;13(11):925-927. doi: 10.1038/nmeth.4004. Epub 2016 Sep 26.

Enzyme-catalyzed expressed protein ligation.

Author information

1
Department of Pharmacology and Molecular and Molecular Sciences, Johns Hopkins School of Medicine, Baltimore, MD, USA.
2
Department of Pharmaceutical Chemistry and Department of Cellular & Molecular Pharmacology, University of California, San Francisco, San Francisco, CA, USA.
#
Contributed equally

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.

PMID:
27669326
PMCID:
PMC5088058
DOI:
10.1038/nmeth.4004
[Indexed for MEDLINE]
Free PMC Article

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