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. 2021 Apr 14;22(6):e50684. doi: 10.15252/embr.202050684

Figure 2. The N‐terminus of USP36 is necessary for promoting SUMOylation.

Figure 2

  1. H1299 cells were transfected with Flag‐USP36 or the indicated deletion mutants and assayed by IB. FL, full length.
  2. Schematic diagram of USP36 indicating that the N‐terminus of USP36, which contains the USP catalytic domain, is required for its SUMO E3 activity. Catalytic residues C131 and H382 are shown.
  3. H1299 cells transfected with WT USP36, the C131A, or H382A mutant were assayed by IB to detect SUMOylated proteins.
  4. H1299 cells transfected with WT USP36 or the indicated mutants and V5‐SUMO2 were assayed by IB using anti‐V5 antibody to detect total protein SUMOylation.
  5. H1299 cells transfected with WT USP36 or the indicated mutants and His‐SUMO2 were subjected to Ni2+‐NTA agarose beads pulldown (PD) followed by IB using anti‐SUMO2/3 antibody.

Source data are available online for this figure.