App Note I November 12, 2018

Sizing Analysis of the Ubiquitin / E1-E2-E3 Complex using switchSENSE® technology 

During the posttranslational process of ubiquitination, the small regulatory protein ubiquitin (Ub) is covalently attached to a lysine residue of a substrate protein in an enzymatic cascade to label target substrates for proteasomal degradation. Alterations in the ubiquitination pathway have been implicated in several diseases, such as in neurodegenerative diseases (e.g. Parkinson’s diseases) as well as in a variety of different types of cancers.

Here, we report a novel approach to study the charge reaction of ubiquitin to the ubiquitin-conjugating enzyme (E2) mediated by the ubiquitin-activating enzyme (E1), as well as the subsequent transfer of ubiquitin to a lysine in a ubiquitin ligase (E3)-dependent discharge reaction using switchSENSE® size analysis.