Highly Selective Lysine Acylation in Proteins Using a Lys‐His Tag Sequence. Issue 15 (17th February 2022)

• Record Type: Journal Article
• Authors: Kofoed, Christian; Wu, Shunliang; Sørensen, Kasper K.; Treiberg, Tuule; Arnsdorf, Johnny; Bjørn, Sara P.; Jensen, Tanja L.; Voldborg, Bjørn G.; Thygesen, Mikkel B.; Jensen, Knud J.; Schoffelen, Sanne
• Is Part Of: Chemistry. Volume 28:Issue 15(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-02-17
• Subjects: acylation -- antibodies -- chemical biology -- peptide tags -- site-selective protein modification; Chemistry -- Periodicals; 540
• DOI: 10.1002/chem.202200147
• Languages: English
• ISSNs: 0947-6539

Abstract

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 N α ‐amine resulting in stable conjugates. Herein, we report the peptide sequences His n ‐Lys‐His m, which we term Lys‐His tags. In combination with simple acylating agents, they facilitate the acylation of the designated Lys N ϵ ‐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. Abstract : A short, designed Lys‐His tag can be inserted into proteins where it autocatalyzes the highly selective acylation of the Lys side‐chain amine with phenyl esters. The reaction conditions of this site‐selective, chemical protein modification method are mild, and no oxidative reagents, reductants nor enzymes are required.