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- W2045012604 abstract "Significance The identification of specific ubiquitin ligase–substrate pairs is crucial for understanding the roles of protein ubiquitination in the regulation of diverse biological processes. Despite the development of various methodologies for substrate identification, it remains challenging to determine ubiquitin ligase substrates. Based on previously described tandem ubiquitin-binding entity(ies) (TUBE), we designed the trypsin-resistant (TR)-TUBE for expression in cells. The coexpression of TR-TUBE with an ubiquitin ligase stabilizes the ubiquitinated substrates by masking the ubiquitin chains. Using a combination of two strategies for enriching ubiquitinated substrates, TR-TUBE and anti–Lys-ε-Gly-Gly antibody, we successfully identified specific ubiquitin ligase–substrate pairs. Our methodology provides an effective means for the identification of ubiquitin ligase substrates and the detection of ubiquitin ligase activity." @default.
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- W2045012604 date "2015-03-31" @default.
- W2045012604 modified "2023-10-18" @default.
- W2045012604 title "A comprehensive method for detecting ubiquitinated substrates using TR-TUBE" @default.
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- W2045012604 doi "https://doi.org/10.1073/pnas.1422313112" @default.
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