1. Sequential release of interacting proteins and Ub-modifying enzymes by disulfide heterotypic ubiquitin reagents.
- Author
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Cai, Hongyi, Wu, Xiangwei, Mao, Junxiong, Tong, Zebin, Yan, Dingfei, Weng, Yicheng, and Zheng, Qingyun
- Subjects
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UBIQUITIN , *ENZYMES , *PROTEINS , *PROTEOMICS - Abstract
• Heterotypic Ub reagents were obtained by CAET strategy with high efficiency. • The reagents identify heterotypic Ub-interacting and modifying proteins simultaneously. • Sequential release of two groups of proteins was completed by using urea and reductant. • Proteomic profiling facilitates future investigations of heterotypic Ub chains. Heterotypic ubiquitin (Ub) chains have emerged as fundamental components in a wide range of cellular processes. The integrative identification of Ub-interacting proteins (readers) and Ub-modifying enzymes (writers and erasers) that selectively recognize and regulate heterotypic ubiquitination may provide crucial insights into these processes. In this study, we employed the bifunctional molecule-assisted (CAET) strategy to develop a type of disulfide bond-activated heterotypic Ub reagents, which allowed to enrich heterotypic Ub-interacting proteins and modifying enzymes simultaneously. The sequential release of readers which are non-covalently bound and writers or erasers which are covalently conjugated by using urea and reductant, respectively, combined with label-free quantitative (LFQ) MS indicated that these heterotypic Ub reagents would facilitate future investigations into functional roles played by heterotypic Ub chains. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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