Proteomics

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Proteomic analysis of linear ubiquitination substrates based on NAEK-Ub probe in live cells

ABSTRACT: While linear ubiquitin plays critical roles in multiple cell signaling pathways, few substrates have been identified. Global profiling of linear ubiquitin substrates represents a significant challenge because of the low endogenous level of linear ubiquitination, the none-specific binding of linear polyubiquitin chain, and the high background interferences arising from heterogeneous ubiquitin linkages (e.g. K48-, K63-). We developed a bioorthogonal linear ubiquitin probe by site-specific encoding of a norbornene amino acid (NAEK) on ubiquitin. This probe facilitates covalent labeling of linear ubiquitin substrates in live cells and enables selective enrichment and identification of linear-ubiquitin modified proteins. we applied this probe in the profiling of substrates of linear ubiquitination in HEK293T cells and GSCs.

ORGANISM(S): Homo Sapiens

SUBMITTER: Xiaotao Duan  

PROVIDER: PXD031600 | iProX | Fri Feb 11 00:00:00 GMT 2022

REPOSITORIES: iProX

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Publications

A bio-orthogonal linear ubiquitin probe identifies STAT3 as a direct substrate of OTULIN in glioblastoma.

Du Xianli X   Pang Jing J   Gu Bin B   Si Tian T   Chang Yan Y   Li Tianqi T   Wu Min M   Wang Zicheng Z   Wang Yuxia Y   Feng Jiannan J   Wu Ning N   Man Jianghong J   Li Huiyan H   Li Ailing A   Zhang Tong T   Wang Bo B   Duan Xiaotao X  

Nucleic acids research 20230201 3

While linear ubiquitin plays critical roles in multiple cell signaling pathways, few substrates have been identified. Global profiling of linear ubiquitin substrates represents a significant challenge because of the low endogenous level of linear ubiquitination and the background interference arising from highly abundant ubiquitin linkages (e.g. K48- and K63-) and from the non-specific attachment of interfering proteins to the linear polyubiquitin chain. We developed a bio-orthogonal linear ubiq  ...[more]

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