TIPI

• Verfasst von: Taxis, Christof [VerfasserIn]
• Verfasst von: Knop, Michael [VerfasserIn]
• Titel: TIPI
• Titelzusatz: TEV protease-mediated induction of protein instability
• Verf.angabe: Christof Taxis, Michael Knop
• Umfang: 16 S.
• Fussnoten: Gesehen am 10.08.2017
• Titel Quelle: Enthalten in: Ubiquitin family modifiers and the proteasome
• Jahr Quelle: 2012
• Band/Heft Quelle: (2012), S. 611-626
• ISBN Quelle: 978-1-61779-474-2
• DOI: doi:10.1007/978-1-61779-474-2_43
• Datenträger: Online-Ressource
• Sprache: eng
• K10plus-PPN: 1562319442

Abstract

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.