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Convenient site-selective protein coupling from bacterial raw lysates to coenzyme A-modified tobacco mosaic virus (TMV) by Bacillus subtilis Sfp phosphopantetheinyl transferase

A facile enzyme-mediated strategy enables site-specific covalent one-step coupling of genetically tagged luciferase molecules to coenzyme A-modified tobacco mosaic virus (TMV-CoA) both in solution and on solid supports. Bacillus subtilis surfactin phosphopantetheinyl transferase Sfp produced in E. c...

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Bibliographische Detailangaben
Hauptverfasser: Geiger, Fania (VerfasserIn) , Wendlandt, Tim (VerfasserIn) , Berking, Tim (VerfasserIn) , Spatz, Joachim P. (VerfasserIn) , Wege, Christina (VerfasserIn)
Dokumenttyp: Article (Journal)
Sprache:Englisch
Veröffentlicht: January 2023
In: Virology
Year: 2023, Jahrgang: 578, Pages: 61-70
ISSN:1096-0341
DOI:10.1016/j.virol.2022.11.013
Online-Zugang:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1016/j.virol.2022.11.013
Verlag, lizenzpflichtig, Volltext: https://www.sciencedirect.com/science/article/pii/S0042682222002112
Volltext
Verfasserangaben:Fania Geiger, Tim Wendlandt, Tim Berking, Joachim P. Spatz, Christina Wege
Beschreibung
Zusammenfassung:A facile enzyme-mediated strategy enables site-specific covalent one-step coupling of genetically tagged luciferase molecules to coenzyme A-modified tobacco mosaic virus (TMV-CoA) both in solution and on solid supports. Bacillus subtilis surfactin phosphopantetheinyl transferase Sfp produced in E. coli mediated the conjugation of firefly luciferase N-terminally extended by eleven amino acids forming a ‘ybbR tag’ as Sfp-selective substrate, which even worked in bacterial raw lysates. The enzymes displayed on the protein coat of the TMV nanocarriers exhibited high activity. As TMV has proven a beneficial high surface-area adapter template stabilizing enzymes in different biosensing layouts in recent years, the use of TMV-CoA for fishing ybbR-tagged proteins from complex mixtures might become an advantageous concept for the versatile equipment of miniaturized devices with biologically active proteins. It comes along with new opportunities for immobilizing multiple functionalities on TMV adapter coatings, as desired, e.g., in handheld systems for point-of-care detection.
Beschreibung:Online verfügbar 26 November 2022, Artikelversion 3 Dezember 2022
Gesehen am 30.01.2023
Beschreibung:Online Resource
ISSN:1096-0341
DOI:10.1016/j.virol.2022.11.013

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