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
Download
![Open access file OA](https://pub.uni-bielefeld.de/images/access_open.png)
Autor*in
Einrichtung
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
Urheberrecht / Lizenzen
eISSN
1439-7633
Finanzierungs-Informationen
Open-Access-Publikationskosten wurden durch die Universität Bielefeld im Rahmen des DEAL-Vertrags gefördert.
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. https://doi.org/10.1002/cbic.202000416
Janson, Nils, Krüger, Tobias, Karsten, Lennard, Boschanski, Mareile, Dierks, Thomas, Müller, Kristian, and Sewald, Norbert. 2020. “Bifunctional Reagents for Formylglycine Conjugation: Pitfalls and Breakthroughs”. ChemBioChem 21 (24): 3580-3593.
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.
Alle Dateien verfügbar unter der/den folgenden Lizenz(en):
Creative Commons Namensnennung - Nicht-kommerziell - Keine Bearbeitung 3.0 Unported (CC BY-NC-ND 3.0):
Volltext(e)
Name
cbic.202000416-2.sewald.pdf
6.14 MB
Access Level
![Open access file OA](https://pub.uni-bielefeld.de/images/access_open.png)
Zuletzt Hochgeladen
2020-12-17T08:23:29Z
MD5 Prüfsumme
05c5414472e5dccd597d791e3fc0be93
Daten bereitgestellt von European Bioinformatics Institute (EBI)
Zitationen in Europe PMC
Daten bereitgestellt von Europe PubMed Central.
References
Daten bereitgestellt von Europe PubMed Central.
Export
Markieren/ Markierung löschen
Markierte Publikationen
Web of Science
Dieser Datensatz im Web of Science®Quellen
PMID: 32767537
PubMed | Europe PMC
Suchen in