TIPI: TEV protease-mediated induction of protein instability

Reverse genetics approaches require methods to inactivate a specific protein. One possibility is to modify the target protein with a degradation signal (degron). Degrons are short, transferable sequences that confer protein instability. They target proteins for degradation either constitutively or a...

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Hauptverfasser: Taxis, Christof (VerfasserIn) , Knop, Michael (VerfasserIn)
Dokumenttyp: Kapitel/Artikel
Sprache:Englisch
Veröffentlicht: 11 January 2012
In: Ubiquitin family modifiers and the proteasome
Year: 2012, Pages: 611-626
DOI:10.1007/978-1-61779-474-2_43
Online-Zugang:Verlag, Volltext: http://dx.doi.org/10.1007/978-1-61779-474-2_43
Verlag, Volltext: https://link.springer.com/protocol/10.1007/978-1-61779-474-2_43
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Verfasserangaben:Christof Taxis, Michael Knop
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Zusammenfassung:Reverse genetics approaches require methods to inactivate a specific protein. One possibility is to modify the target protein with a degradation signal (degron). Degrons are short, transferable sequences that confer protein instability. They target proteins for degradation either constitutively or after activation, e.g., by phosphorylation, presence of a binding partner, or conformational rearrangements in the substrate. In this chapter, we describe a synthetic way to activate a degron. It employs the generation of an N-degron by cleavage of a substrate with the site-specific tobacco etch virus (TEV) protease. Subsequently, the substrate is targeted for degradation by the ubiquitin-proteasome system. This TEV protease-induced protein instability system provides a powerful approach to generate conditional mutants for synthetic biology or for the investigation of protein functions in a specific cellular context.
Beschreibung:Gesehen am 10.08.2017
Beschreibung:Online Resource
ISBN:9781617794742
DOI:10.1007/978-1-61779-474-2_43