Comprehensive Identification of Substrates for F-box Proteins by Differential Proteomics Analysis
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- 作者：Kanae Yumimoto ; Masaki Matsumoto ; Koji Oyamada ; Toshiro Moroishi ; Keiichi I. Nakayama
- 刊名：Journal of Proteome Research
- 出版年：2012
- 出版时间：June 1, 2012
- 年：2012
- 卷：11
- 期：6
- 页码：3175-3185
- 全文大小：756K
- 年卷期：v.11,no.6(June 1, 2012)
- ISSN：1535-3907
文摘
Although elucidation of enzyme鈥搒ubstrate relations is fundamental to the advancement of biology, universal approaches to the identification of substrates for a given enzyme have not been established. It is especially difficult to identify substrates for ubiquitin ligases, given that most such substrates are immediately ubiquitylated and degraded as a result of their association with the enzyme. We here describe the development of a new approach, DiPIUS (differential proteomics-based identification of ubiquitylation substrates), to the discovery of substrates for ubiquitin ligases. We applied DiPIUS to Fbxw7伪, Skp2, and Fbxl5, three of the most well-characterized F-box proteins, and identified candidate substrates including previously known targets. DiPIUS is thus a powerful tool for unbiased and comprehensive screening for substrates of ubiquitin ligases.