| Literature DB >> 28319114 |
Katarzyna Kliza1, Christoph Taumer2, Irene Pinzuti3, Mirita Franz-Wachtel2, Simone Kunzelmann4, Benjamin Stieglitz3, Boris Macek2, Koraljka Husnjak1.
Abstract
Ubiquitination controls a plethora of cellular processes. Modifications by linear polyubiquitin have so far been linked with acquired and innate immunity, lymphocyte development and genotoxic stress response. Until now, a single E3 ligase complex (LUBAC), one specific deubiquitinase (OTULIN) and a very few linear polyubiquitinated substrates have been identified. Current methods for studying lysine-based polyubiquitination are not suitable for the detection of linear polyubiquitin-modified proteins. Here, we present an approach to discovering linear polyubiquitin-modified substrates by combining a lysine-less internally tagged ubiquitin (INT-Ub.7KR) with SILAC-based mass spectrometry. We applied our approach in TNFα-stimulated T-REx HEK293T cells and validated several newly identified linear polyubiquitin targets. We demonstrated that linear polyubiquitination of the novel LUBAC substrate TRAF6 is essential for NFκB signaling.Entities:
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Year: 2017 PMID： 28319114 DOI： 10.1038/nmeth.4228
Source DB: PubMed Journal: Nat Methods ISSN： 1548-7091 Impact factor: 28.547