TIPI: TEV protease-mediated induction of protein instability.
TIPI: TEV protease-mediated induction of protein instability.
复制标题
TIPI：TEV 蛋白酶介导的蛋白质不稳定诱导。
作者：
Christof Taxis;M. Knop
反向遗传学方法需要对特定蛋白质进行测序的方法。一种可能性是用降解信号（降解决定子）修饰靶蛋白。降解决定子是赋予蛋白质不稳定性的短的、可转移的序列。它们靶向蛋白质以进行组成型降解或活化后降解，例如，通过磷酸化、结合伴侣的存在或底物中的构象重排。在本章中，我们描述了一种激活降解决定子的合成方法。它采用通过用位点特异性烟草蚀纹病毒（TEV）蛋白酶切割底物来产生N-降解决定子。随后，底物被泛素-蛋白酶体系统靶向降解。这种TEV蛋白酶诱导的蛋白质不稳定性系统提供了一个强大的方法来产生条件突变体的合成生物学或在特定的细胞环境中的蛋白质功能的调查。
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.
影响因子：
56.9
作者：
BACHMAIR, A;FINLEY, D;VARSHAVSKY, A
通讯作者：
VARSHAVSKY, A