Fig. 1: The principle of the neutron-encoded diUb cleavage assay.
From: Neutron-encoded diubiquitins to profile linkage selectivity of deubiquitinating enzymes
Top—Schematic outline of classical diUb cleavage assays relying on separate incubation of each diUb isoform with a DUB followed by SDS-PAGE, MS or fluorescent intensity read-out for every diUb isoform separately. Bottom—Schematic outline of the designed method where a DUB is incubated with a mixture containing all neutron-encoded diUb isoforms followed by MS analysis, allowing the quantification of each linkage from the complex mixture. Upon cleavage of the diUb by a DUB, the diUb signal(s) will disappear and the corresponding monoUb signal(s) will appear in the MS spectrum. Colors represent the differently linked diUbs as indicated.
