Sequential release of interacting proteins and Ub-modifying enzymes by disulfide heterotypic ubiquitin reagents

ABSTRACT: Heterotypic ubiquitin (Ub) chains have emerged as fundamental components in a wide range of cellular processes. The integrative identification of Ub-interacting proteins (readers) and Ub-modifying enzymes (writers and erasers) that selectively recognize and regulate heterotypic ubiquitination may provide crucial insights into these processes. In this study, we employed the bifunctional molecule-assisted (CAET) strategy to develop a new type of disulfide-bond-activated heterotypic Ub reagents, which allowed to enrich heterotypic Ub-interacting proteins and modifying enzymes simultaneously. The sequential release of readers which are non-covalently bound and writers or erasers which are covalently conjugated by using urea and reductant, respectively, combined with label-free quantitative (LFQ) MS indicated that these innovative heterotypic Ub tools would facilitate future investigations into functional roles played by heterotypic Ub chains.

ORGANISM(S): Saccharomyces Cerevisiae

SUBMITTER: Qingyun Zheng  

PROVIDER: PXD045936 | iProX | Thu Sep 28 00:00:00 BST 2023

REPOSITORIES: iProX