Highly Selective Lysine Acylation in Proteins Using a Lys-His Tag Sequence |
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Authors: | Dr. Christian Kofoed Dr. Shunliang Wu Dr. Kasper K. Sørensen Tuule Treiberg Dr. Johnny Arnsdorf Dr. Sara P. Bjørn Dr. Tanja L. Jensen Bjørn G. Voldborg Dr. Mikkel B. Thygesen Prof. Knud J. Jensen Dr. Sanne Schoffelen |
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Affiliation: | 1. Department of Chemistry, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg, Denmark;2. National Biologics Facility, Department of Biotechnology and Biomedicine, Technical University of Denmark, Building 220, Kemitorvet, 2800 Kgs. Lyngby, Denmark;3. The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800 Kgs. Lyngby, Denmark |
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Abstract: | 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. Herein, 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. |
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Keywords: | acylation antibodies chemical biology peptide tags site-selective protein modification |
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