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