Two-step protein labeling utilizing lipoic acid ligase and sonogashira cross-coupling

• Verfasst von: Hauke, Sebastian [VerfasserIn]
• Verfasst von: Best, Marcel [VerfasserIn]
• Verfasst von: Schmidt, Tobias Thomas [VerfasserIn]
• Verfasst von: Baalmann, Mathis [VerfasserIn]
• Verfasst von: Krause, André [VerfasserIn]
• Verfasst von: Wombacher, Richard [VerfasserIn]
• Titel: Two-step protein labeling utilizing lipoic acid ligase and sonogashira cross-coupling
• Verf.angabe: Sebastian Hauke, Marcel Best, Tobias T. Schmidt, Mathis Baalmann, André Krause, and Richard Wombacher
• E-Jahr: 2014
• Jahr: August 24, 2014
• Umfang: 6 S.
• Illustrationen: Illustrationen
• Fussnoten: Gesehen am 04.09.2020
• Titel Quelle: Enthalten in: Bioconjugate chemistry
• Ort Quelle: Columbus, Ohio : American Chemical Society, 1990
• Jahr Quelle: 2014
• Band/Heft Quelle: 25(2014), 9, Seite 1632-1637
• ISSN Quelle: 1520-4812
• DOI: doi:10.1021/bc500349h
• Datenträger: Online-Ressource
• Sprache: eng
• K10plus-PPN: 1728906121

Abstract

Labeling proteins in their natural settings with fluorescent proteins or protein tags often leads to problems. Despite the high specificity, these methods influence the natural functions due to the rather large size of the proteins used. Here we present a two-step labeling procedure for the attachment of various fluorescent probes to a small peptide sequence (13 amino acids) using enzyme-mediated peptide labeling in combination with palladium-catalyzed Sonogashira cross-coupling. We identified p-iodophenyl derivatives from a small library that can be covalently attached to a lysine residue within a specific 13-amino-acid peptide sequence by Escherichia coli lipoic acid ligase A (LplA). The derivatization with p-iodophenyl subsequently served as a reactive handle for bioorthogonal transition metal-catalyzed Sonogashira cross-coupling with alkyne-functionalized fluorophores on both the peptide as well as on the protein level. Our two-step labeling strategy combines high selectivity of enzyme-mediated labeling with the chemoselectivity of palladium-catalyzed Sonogashira cross-coupling.