Orthogonal dual-modification of proteins for the engineering of multivalent protein scaffolds

M. Mühlberg, M. G. Hoesl, C. Kuehne, J. Dernedde, N. Budisa, C. P. R. Hackenberger – 2015

To add new tools to the repertoire of protein-based multivalent scaffold design, we have developed a novel dual-labeling strategy for proteins that combines residue-specific incorporation of unnatural amino acids with chemical oxidative aldehyde formation at the N-terminus of a protein. Our approach relies on the selective introduction of two different functional moieties in a protein by mutually orthogonal copper-catalyzed azide–alkyne cycloaddition (CuAAC) and oxime ligation. This method was applied to the conjugation of biotin and β-linked galactose residues to yield an enzymatically active thermophilic lipase, which revealed specific binding to Erythrina cristagalli lectin by SPR binding studies.

Titel
Orthogonal dual-modification of proteins for the engineering of multivalent protein scaffolds
Verfasser
M. Mühlberg, M. G. Hoesl, C. Kuehne, J. Dernedde, N. Budisa, C. P. R. Hackenberger
Datum
2015
Kennung
10.3762/bjoc.11.88
Zitierweise
Beilstein J. Org. Chem. 2015, 11, 784–791
Art
Text
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