Two‐fold Bioorthogonal Derivatization by Different Formylglycine‐Generating Enzymes

Formylglycine‐generating enzymes are of increasing interest in the field of bioconjugation chemistry. They catalyze the site‐specific oxidation of a cysteine residue to the aldehyde‐containing amino acid Cα‐formylglycine (FGly). This non‐canonical residue can be generated within any desired target p...

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Bibliographic Details
Published in:Angewandte Chemie International Edition 2018-06, Vol.57 (24), p.7245-7249
Main Authors: Krüger, Tobias, Weiland, Stefanie, Falck, Georg, Gerlach, Marcus, Boschanski, Mareile, Alam, Sarfaraz, Müller, Kristian M., Dierks, Thomas, Sewald, Norbert
Format: Article
Language:English
Subjects:
Amino acids
Bacteria - enzymology
bioconjugation
Cell Line
Conjugation
Cysteine - metabolism
enzyme catalysis
Enzymes
formylglycine
Glycine - analogs & derivatives
Glycine - metabolism
Humans
Iron
Iron-Sulfur Proteins - metabolism
Methanosarcina - enzymology
Organic chemistry
Oxidation
Oxidation-Reduction
Peptides
Proteins
radical-SAM enzymes
Sulfatases - metabolism
Sulfur
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Summary:Formylglycine‐generating enzymes are of increasing interest in the field of bioconjugation chemistry. They catalyze the site‐specific oxidation of a cysteine residue to the aldehyde‐containing amino acid Cα‐formylglycine (FGly). This non‐canonical residue can be generated within any desired target protein and can subsequently be used for bioorthogonal conjugation reactions. The prototypic formylglycine‐generating enzyme (FGE) and the iron‐sulfur protein AtsB display slight variations in their recognition sequences. We designed specific tags in peptides and proteins that were selectively converted by the different enzymes. Combination of the different tag motifs within a single peptide or recombinant protein enabled the independent and consecutive introduction of two formylglycine residues and the generation of heterobifunctionalized protein conjugates. One Way or Another: The formylglycine (FGly)‐based bioconjugation strategy was expanded by utilizing the iron‐sulfur protein AtsB. By combining AtsB with the prototypic formylglycine generating enzyme (FGE), the independent introduction of two aldehyde moieties into peptides and proteins was accomplished. This bioorthogonal enzymatic system represents a new strategy for dual protein labeling at two specific sites.
ISSN:1433-7851
1521-3773
DOI:10.1002/anie.201803183