Middle-down mass spectrometry enables characterization of branched ubiquitin chains.

ABSTRACT: Protein ubiquitylation, one of the most prevalent post-translational modifications in eukaryotes, is involved in regulating nearly every cellular signaling pathway. The vast functional range of ubiquitylation has largely been attributed to the formation of a diverse array of polymeric ubiquitin (polyUb) chains. Methods that enable the characterization of these diverse chains are necessary to fully understand how differences in structure relate to function. Here, we describe a method for the detection of enzymatically derived branched polyUb conjugates in which a single Ub subunit is modified by two Ub molecules at distinct lysine residues. Using a middle-down mass spectrometry approach in which restricted trypsin-mediated digestion is coupled with mass spectrometric analysis, we characteri

SUBMITTER: Valkevich EM 

PROVIDER: S-EPMC4372068 | biostudies-literature | 2014 Aug

REPOSITORIES: biostudies-literature