Advances in characterizing ubiquitylation sites by mass spectrometry

• Published in: Current opinion in chemical biology Vol. 17; no. 1; pp. 49 - 58
• Main Authors: Sylvestersen, Kathrine B, Young, Clifford, Nielsen, Michael L
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.02.2013
• Subjects: Amino Acid Sequence; Animals; Humans; Mass Spectrometry - methods; Molecular Sequence Data; Ubiquitin - chemistry; Ubiquitin - metabolism; Ubiquitinated Proteins - chemistry; Ubiquitinated Proteins - metabolism; Ubiquitination
• ISSN: 1367-5931; 1879-0402
• DOI: 10.1016/j.cbpa.2012.12.009

► Ubiquitin is a widely occurring and dynamic post-translational modification. ► Mass spectrometry has emerged as a powerful methodology to characterize ubiquitylation sites. ► Peptide-centric enrichment strategies are currently superior to protein-centric methodologies. The attachment of one or more ubiquitin moieties to proteins plays a central regulatory mechanism in eukaryotic cells. Protein ubiquitylation regulates numerous cellular processes, including protein degradation, signal transduction, DNA repair and cell division. The characterization of ubiquitylation is a two-fold challenge that involves the mapping of ubiquitylation sites and the determination of ubiquitin chain topology. This review focuses on the technical advances in the mass spectrometry-based characterization of ubiquitylation sites, which have recently involved the large-scale identification of ubiquitylation sites by peptide-level enrichment strategies. The discovery that ubiquitylation is a widespread modification similar to phosphorylation and acetylation suggests cross-talk may also occur at the post translational modification level.