Preparation, characterization, and use of tagged ubiquitins

被引：8

作者：

Callis, J ^{[1
]}

Ling, R ^{[1
]}

机构：

[1] Univ Calif Davis, Sect Mol & Cellular Biol, Davis, CA 95616 USA

来源：

UBIQUITIN AND PROTEIN DEGRADATION, PT B | 2005年 / 399卷

基金：

美国国家科学基金会;

关键词：

D O I：

10.1016/S0076-6879(05)99004-6

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The low abundance and heterogeneity of ubiquitinated proteins has led to the development and use of tagged forms of ubiquitin. Additional residues present at the ubiquitin amino terminus provide immunological and/or affinity sites to facilitate visualization, identification, and purification of ubiquitinated substrates by virtue of their covalent attachment to the tagged ubiquitin. The use of tagged ubiquitin to understand the scope, nature, and biological relevance of this conserved modification system has been demonstrated in multiple ways. Unknown substrates can be identified, or a previously identified substrate can be analyzed with tagged ubiquitin in vitro or in vivo to determine the specificity, regulation, and type of ubiquitin linkages formed. This contribution describes the generation and use of multiple types of modified ubiquitins: biotinylated ubiquitin produced in vitro, or GST-, myc-, HA-, and hexahistidine-tagged ubiquitins produced in vivo.

引用

页码：51 / 64

页数：14

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