Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 54, Issue 2, Pages 441-445Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201408126
Keywords
biotechnology; protein modification; bioconjugation; sortase
Categories
Funding
- National Institutes of Health [R01 GM061232]
- University Scholars Fellowship - Duke University Graduate School
- NIH [5T32 GM008487]
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We provide the first demonstration that isopeptide ligation, a noncanonical activity of the enzyme sortaseA, can be used to modify recombinant proteins. This reaction was used invitro to conjugate small molecules to a peptide, an engineered targeting protein, and a full-length monoclonal antibody with an exquisite level of control over the site of conjugation. Attachment to the protein substrate occurred exclusively through isopeptide bonds at a lysine epsilon-amino group within a specific amino acid sequence. This reaction allows more than one molecule to be site-specifically conjugated to a protein at internal sites, thereby overcoming significant limitations of the canonical native peptide ligation reaction catalyzed by sortaseA. Our method provides a unique chemical ligation procedure that is orthogonal to existing methods, supplying a new method to site-specifically modify lysine residues that will be a valuable addition to the protein conjugation toolbox.
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