Extended Data Fig. 1: SidJ does not possess intrinsic deubiquitinase activity.
From: Inhibition of bacterial ubiquitin ligases by SidJ–calmodulin catalysed glutamylation
a, Genetic locus of sdeC-orf2-sidj-sdeB-sdeA in the Legionella genome. b, Left, GFP–SidJ that was ectopically expressed and purified from HEK293T cells was incubated with canonical HA–ubiquitin chains purified from mammalian cells. The canonical deubiquitinase USP2 was used as a positive control. Right, GFP or GFP–SidJ was incubated with purified SdeA–ubiquitinated Rab33b. The experiment was repeated twice independently, with similar results. c, Full-length SidJ was incubated with various substrates modified with canonical ubiquitination or phosphoribosyl-linked ubiquitination, to probe the cleavage activity. USP2 was used as a positive control for cleaving canonical ubiquitin chains. The experiment was repeated twice independently, with similar results. d, SidJ purified from Legionella was analysed by mass spectrometry, and protein quantification was performed using the MaxQuant iBAQ algorithm. The experiment was repeated twice independently, with similar results.
