Journal article
Highly selective lysine acylation in proteins using a Lys-His tag sequence
University of Copenhagen1
Department of Biotechnology and Biomedicine, Technical University of Denmark2
Section for Protein Science and Biotherapeutics, Department of Biotechnology and Biomedicine, Technical University of Denmark3
National Biologics Facility, Section for Protein Science and Biotherapeutics, Department of Biotechnology and Biomedicine, Technical University of Denmark4
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark5
Chemical modification of proteins has numerous applications, but it has been challenging to achieve the required high degree of selectivity on lysine amino groups. Recently, we described the highly selective acylation of proteins with an N-terminal Gly-His6 segment. This tag promoted acylation of the N-terminal Nα -amine resulting in stable conjugates.
Here, we report the peptide sequences Hisn-Lys-Hism , which we term Lys-His tags. In combination with simple acylating agents, they facilitate the acylation of the designated Lys Nε-amine under mild conditions and with high selectivity over native Lys residues. We show that the Lys-His tags, which are 7 to 10 amino acids in length and still act as conventional His tags, can be inserted in proteins at the C-terminus or in loops, thus providing high flexibility regarding the site of modification.
Finally, the selective and efficient acylation of the therapeutic antibody Rituximab, pure or mixed with other proteins, demonstrates the scope of the Lys-His tag acylation method.
Language: | English |
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Year: | 2022 |
Pages: | e202200147 |
ISSN: | 15213757 , 00448249 , 09476539 and 15213765 |
Types: | Journal article |
DOI: | 10.1002/chem.202200147 |
ORCIDs: | 0000-0002-0158-2802 , 0000-0003-3525-5452 , 0000-0002-8258-5887 , 0000-0002-8590-7124 , 0000-0003-3070-9144 , Arnsdorf, Johnny , Bjørn, Sara P. , Voldborg, Bjørn G. , Schoffelen, Sanne and Jensen, Tanja L. |