A strategy for orthogonal deubiquitination using a bump-and-hole approach

We have successfully applied a bump-and-hole approach to establish orthogonal deubiquitination in which a ubiquitin substrate variant is specifically targeted by an engineered deubiquitinating enzyme (DUB). This makes it possibe to selectively observe and measure a single type of DUB activity in living cells. We established orthogonal deubiquitination, which enables us to selectively observe a single type of deubiquitinating enzyme activity in living cells.