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Nat Commun. 2018 Aug 17;9(1):3307. doi: 10.1038/s41467-018-05695-3.

Selective N-terminal acylation of peptides and proteins with a Gly-His tag sequence.

Author information

1
Department of Chemistry, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg, Denmark.
2
Biomolecular Nanoscale Engineering Center, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg, Denmark.
3
Niels Bohr Institute, University of Copenhagen, Universitetsparken 5, 2100, Copenhagen, Denmark.
4
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet Building 220, 2800, Kgs. Lyngby, Denmark.
5
Department of Chemistry, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg, Denmark. sanne@chem.ku.dk.
6
Center for Evolutionary Chemical Biology, University of Copenhagen, Universitetsparken 5, 2100, Copenhagen, Denmark. sanne@chem.ku.dk.
7
Department of Chemistry, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg, Denmark. kjj@chem.ku.dk.
8
Biomolecular Nanoscale Engineering Center, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg, Denmark. kjj@chem.ku.dk.

Abstract

Methods for site-selective chemistry on proteins are in high demand for the synthesis of chemically modified biopharmaceuticals, as well as for applications in chemical biology, biosensors and more. Inadvertent N-terminal gluconoylation has been reported during expression of proteins with an N-terminal His tag. Here we report the development of this side-reaction into a general method for highly selective N-terminal acylation of proteins to introduce functional groups. We identify an optimized N-terminal sequence, GHHHn- for the reaction with gluconolactone and 4-methoxyphenyl esters as acylating agents, facilitating the introduction of functionalities in a highly selective and efficient manner. Azides, biotin or a fluorophore are introduced at the N-termini of four unrelated proteins by effective and selective acylation with the 4-methoxyphenyl esters. This Gly-Hisn tag adds the unique capability for highly selective N-terminal chemical acylation of expressed proteins. We anticipate that it can find wide application in chemical biology and for biopharmaceuticals.

PMID:
30120230
PMCID:
PMC6098153
DOI:
10.1038/s41467-018-05695-3
[Indexed for MEDLINE]
Free PMC Article

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