Tobias Krüger
Bielefeld University
CytotoxicityConjugated systemInternalizationProtein engineeringPeptide sequenceKnoevenagel condensationEnzymeChemistryConjugateCombinatorial chemistryBioconjugationSerineDARPinFormylglycine-generating enzymeAldehyde tagSodium dodecyl sulfateCysteinePeptideAldehydeTarget proteinBifunctionalMonomethyl auristatin EL-amino-acid oxidaseRadical SAMResidue (chemistry)Immobilized enzymeRecombinant DNAAmino acidConsensus sequenceBiochemistryClick chemistryTurnover numberBioorthogonal chemistry
7Publications
3H-index
46Citations
Publications 7
Newest
#1Mareile Boschanski (Bielefeld University)H-Index: 3
#2Tobias Krüger (Bielefeld University)H-Index: 3
Last. Thomas Dierks (Bielefeld University)H-Index: 45
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Multiple, site-specific protein conjugation is increasingly attractive for the generation of antibody-drug conjugates (ADCs). As it is important to control the number and position of cargoes in an ADC, position-selective generation of reactive sites in the protein of interest is required. Formylglycine (FGly) residues are generated by enzymatic conversion of cysteine residues embedded in a certain amino acid sequence motif with a formylglycine-generating enzyme (FGE). The addition of copper ions...
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#1Nils Janson (Bielefeld University)H-Index: 1
#2Tobias Krüger (Bielefeld University)H-Index: 3
Last. Norbert SewaldH-Index: 52
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Formylglycine-generating enzymes specifically oxidize cysteine within the consensus sequence CxPxR to C α -formylglycine (FGly). This non-canonical electrophilic amino acid can subsequently be addressed selectively via bioorthogonal Hydrazino- iso -Pictet-Spengler (HIPS) or Knoevenagel ligation to attach payloads like fluorophores or drugs to proteins, while obtaining a defined payload-to-protein ratio. However, disadvantages of these conjugation techniques include the need of a large excess of ...
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#1Tobias Krüger (Bielefeld University)H-Index: 3
#2Stefanie Weiland (Bielefeld University)H-Index: 2
Last. Norbert Sewald (Bielefeld University)H-Index: 52
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: Formylglycine-generating enzymes provide a convenient tool for site-specific protein derivatization. Their ability to oxidize cysteine or serine residues within a defined consensus sequence to Cα -formylglycine (FGly) allows for the targeted introduction of a unique chemical handle for various bioconjugation reactions. In recent years, oxygen-dependent FGly-generating enzyme saw broad use in protein functionalization and the generation of protein conjugates. Yet, the FGly-generating system Ats...
1 CitationsSource
#1Tobias Krüger (Bielefeld University)H-Index: 3
#2Thomas Dierks (Bielefeld University)H-Index: 45
Last. Norbert Sewald (Bielefeld University)H-Index: 52
view all 3 authors...
: Site-specific bioconjugation strategies offer many possibilities for directed protein modifications. Among the various enzyme-based conjugation protocols, formylglycine-generating enzymes allow to posttranslationally introduce the amino acid Cα-formylglycine (FGly) into recombinant proteins, starting from cysteine or serine residues within distinct consensus motifs. The aldehyde-bearing FGly-residue displays orthogonal reactivity to all other natural amino acids and can, therefore, be used for...
13 CitationsSource
#1Svenja Bloess (Bielefeld University)H-Index: 2
#2Tobias Beuel (Bielefeld University)H-Index: 1
Last. Gabriele Fischer von Mollard (Bielefeld University)H-Index: 24
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l-Amino acid oxidases (LAAOs) are flavoproteins, which use oxygen to deaminate l-amino acids and produce the corresponding α-keto acids, ammonia, and hydrogen peroxide. Here we describe the heterologous expression of LAAO4 from the fungus Hebeloma cylindrosporum without signal sequence as fusion protein with a 6His tag in Escherichia coli and its purification. 6His-hcLAAO4 could be activated by exposure to acidic pH, the detergent sodium dodecyl sulfate, or freezing. The enzyme converted 14 prot...
12 CitationsSource
#1Tobias Krüger (Bielefeld University)H-Index: 3
#2Stefanie Weiland (Bielefeld University)H-Index: 2
Last. Norbert Sewald (Bielefeld University)H-Index: 52
view all 9 authors...
: 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...
12 CitationsSource
#1Tobias Krüger (Bielefeld University)H-Index: 3
#2Stefanie Weiland (Bielefeld University)H-Index: 2
Last. Norbert Sewald (Bielefeld University)H-Index: 52
view all 9 authors...
2 CitationsSource