Bifunctional Reagents for Formylglycine Conjugation: Pitfalls and Breakthroughs

Janson N, Krüger T, Karsten L, Boschanski M, Dierks T, Müller K, Sewald N (2020)
ChemBioChem 21(24): 3580-3593.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Abstract / Bemerkung
Formylglycine-generating enzymes specifically oxidize cysteine within the consensus sequence CxPxR to C alpha -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 conjugation building block, comparably low reaction rates and limited stability of FGly-containing proteins. Therefore, functionalized clickable HIPS and tandem Knoevenagel building blocks were synthesized, conjugated to small proteins (DARPins) and subsequently linked to strained alkyne-containing payloads for protein labeling. This procedure allowed the selective bioconjugation of one or two DBCO-carrying payloads with nearly stoichiometric amounts at low concentrations. Furthermore, an azide-modified tandem Knoevenagel building block enabled the synthesis of branched PEG-linkers and the conjugation of two fluorophores, resulting in an improved signal-to-noise ratio in live cell fluorescence imaging experiments targeting the EGF-receptor. © 2020 Wiley-VCH GmbH.
Erscheinungsjahr
2020
Zeitschriftentitel
ChemBioChem
Band
21
Ausgabe
24
Seite(n)
3580-3593
eISSN
1439-7633
Page URI
https://pub.uni-bielefeld.de/record/2945531

Zitieren

Janson N, Krüger T, Karsten L, et al. Bifunctional Reagents for Formylglycine Conjugation: Pitfalls and Breakthroughs. ChemBioChem. 2020;21(24):3580-3593.
Janson, N., Krüger, T., Karsten, L., Boschanski, M., Dierks, T., Müller, K., & Sewald, N. (2020). Bifunctional Reagents for Formylglycine Conjugation: Pitfalls and Breakthroughs. ChemBioChem, 21(24), 3580-3593. doi:10.1002/cbic.202000416
Janson, N., Krüger, T., Karsten, L., Boschanski, M., Dierks, T., Müller, K., and Sewald, N. (2020). Bifunctional Reagents for Formylglycine Conjugation: Pitfalls and Breakthroughs. ChemBioChem 21, 3580-3593.
Janson, N., et al., 2020. Bifunctional Reagents for Formylglycine Conjugation: Pitfalls and Breakthroughs. ChemBioChem, 21(24), p 3580-3593.
N. Janson, et al., “Bifunctional Reagents for Formylglycine Conjugation: Pitfalls and Breakthroughs”, ChemBioChem, vol. 21, 2020, pp. 3580-3593.
Janson, N., Krüger, T., Karsten, L., Boschanski, M., Dierks, T., Müller, K., Sewald, N.: Bifunctional Reagents for Formylglycine Conjugation: Pitfalls and Breakthroughs. ChemBioChem. 21, 3580-3593 (2020).
Janson, Nils, Krüger, Tobias, Karsten, Lennard, Boschanski, Mareile, Dierks, Thomas, Müller, Kristian, and Sewald, Norbert. “Bifunctional Reagents for Formylglycine Conjugation: Pitfalls and Breakthroughs”. ChemBioChem 21.24 (2020): 3580-3593.
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2020-12-17T08:23:29Z
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