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Published on: February 5, 2020
Sortase A Catalyzed Transpeptidation for Protein Bioengineering
Floris J van Dalen1, Hidde L Ploegh2
1Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA. Floris.vanDalen@childrens.harvard.edu.
Abstract:
Sortase A, originating from Staphylococcus Aureus, has become the gold standard for site-specific transpeptidation of protein substrates. Wild-type and engineered variants of sortase A catalyze the recognition and cleavage of a five-amino acid substrate motif (LPxTG), followed by transpeptidation with oligoglycine nucleophiles (peptides that contain one or more glycines at their N-terminus). This LPxTG sortagging motif can be introduced by genetic engineering of recombinantly expressed proteins of interest. Oligoglycine nucleophiles can be readily synthesized on solid-phase resin and allow the introduction of a wide range of functional groups. Here, we describe the transpeptidation of two nanobodies (VHHs, variable region of the heavy chain of heavy-chain only antibodies) with various GGG-nucleophiles.
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