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...
Gespeichert in:
| Hauptverfasser: | , |
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| Dokumenttyp: | Kapitel/Artikel |
| Sprache: | Englisch |
| Veröffentlicht: |
11 January 2012
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| 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 |
MARC
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| 520 | |a 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. | ||
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