A diubiquitin-based photoaffinity probe for profiling K27-linkage targeting deubiquitinases
Xiao-Dan Tan1, Man Pan, Shuai Gao
1Tsinghua-Peking Center for Life Sciences, Ministry of Education Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology, Department of Chemistry, Tsinghua University, Beijing 100084, China.
Abstract:
K27-linkage poly-ubiquitination plays important roles in DNA damage repair and autoimmunity. Identification of K27-linkage targeting deubiquitinases (DUBs) is essential for understanding their regulatory mechanisms. Here we report an aryl-azide-based photoaffinity diubiquitin (diUb) probe for profiling K27-linkage targeting DUBs. This probe shows high selectivity and sensitivity towards K27-linkage DUBs in vitro. It can also be used in cell lysate for identifying and probing K27-linkage targeting DUBs in real proteomes. Our work suggests that the photoaffinity-based strategy may provide a new useful approach for the construction of other isopeptide linkage targeting DUB probes.
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