A diubiquitin-based photoaffinity probe for profiling K27-linkage targeting deubiquitinases
A diubiquitin-based photoaffinity probe for profiling K27-linkage targeting deubiquitinases
复制标题
基于双泛素的光亲和探针，用于分析靶向去泛素酶的 K27 连接
DOI：
10.1039/c7cc05504h
复制
发表时间：
2017
影响因子：
4.9
通讯作者：
Li Yi-Ming
中科院分区：
文献类型：
--
作者：
Tan Xiao-Dan;Pan Man;Gao Shuai;Zheng Yong;Shi Jing;Li Yi-Ming
K27连接的多聚泛素化在DNA损伤修复和自身免疫中起重要作用。靶向K27连接的去遍在蛋白酶（DUB）的鉴定对于了解其调节机制至关重要。在这里，我们报告了一个芳基叠氮化物为基础的光亲和双泛素（diUb）探针分析K27连接靶向DUBs。该探针在体外对K27-连接DUBs显示出高选择性和灵敏度。它还可用于细胞裂解物中，以识别和探测真实的蛋白质组中靶向K27连接的DUB。我们的工作表明，基于光亲和性的策略可能会提供一个新的有用的方法来构建其他异肽键靶向DUB探针。
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.