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. 2014 Dec 20;34(3):307–325. doi: 10.15252/embj.201489847

Figure 8.

Figure 8

Defects in hydrolyzing phosphoUb chains
  • A Structure of USP2 (red) bound to Ub (cyan) [pdb-id 2hd5 (Renatus et al, 2006)], with a close-up on Ub Ser65 in the distal Ub. A negative charge at the USP2 Asp345 position is conserved in most USP enzymes (Ye et al, 2009).
  • B-H Ubiquitination reactions from Fig6A were used as input for deubiquitinase (DUB) analysis. Silver-stained time-course gels are shown. All DUBs tested (USP2, USP8, USP15, USP30, Ataxin-3, USP21) hydrolyze Ub chains but have significantly lower activity against phosphoUb chains, with the exception of vOTU and USP21.
  • I USP30 activity was significantly lower for cIAP polyUb as compared to other USP DUBs. Cleavage assays against the diUb panel reveals preference for Lys6-linked chains.
  • J USP30 activity is decreased against phosphorylated Lys6 diUb.
  • K-N Deubiquitinase assays against phosphorylated tetraUb. The activity of AMSH (K), TRABID (L), OTUB1 (M) and OTULIN (N) are inhibited with Lys63, Lys33, Lys48 and Met1 phosphorylated tetraUb, respectively.