泛素连接酶
泛素
泛素蛋白连接酶类
调节器
细胞生物学
泛素结合酶
F盒蛋白
化学
生物化学
生物
基因
作者
Kazuhiro Iwaï,Haruto Ishikawa,Takayoshi Kirisako
出处
期刊:Methods in Enzymology
日期:2005-01-01
卷期号:: 256-271
被引量:5
标识
DOI:10.1016/s0076-6879(05)98021-x
摘要
The ubiquitin system plays important roles in the regulation of numerous cellular processes. It is well established that ubiquitin ligases (E3s) are key components in determining the specificity of the system and that the modification of substrates such as phosphorylation often plays a critical role in selective substrate recognition by E3s. Through studies analyzing iron‐mediated degradation of iron regulatory protein 2 (IRP2), a central regulator of iron metabolism in mammalian cells, we have identified a RING finger protein, HOIL‐1, as an ubiquitin ligase recognizing IRP2 through a signal created by heme‐mediated oxidative modification of the protein. We have utilized several types of in vitro ubiquitination assays that detect IRP2 ubiquitination and a differential yeast two‐hybrid screen in which yeast cells were cultured either in the presence or in the absence of oxygen to control the oxidation state of the bait in the cells in our studies. This chapter describes the detailed methods used for the identification and functional analysis of the HOIL‐1 ligase.
科研通智能强力驱动
Strongly Powered by AbleSci AI