Fig. 4

ZUFSP UBDs contribute to chain cleavage and specificity. a Schematic representation of ZUFSP domain architecture. UBZ-like zinc fingers (Z), MIU domain (M), novel ZUFSP ubiquitin-binding domain (zUBD) and α2/α3 region are shown as boxes. b Activity of ZUFSP FL and truncations lacking the UBDs against K63-linked Ub₆₊ chains. Positions of the truncated ZUFSP proteins are indicated by arrows. c Comparison of Mug105 and ZUFSP^310-578 activity against ubiquitin-AMC. The shown RFU values are the mean of triplicates. d Chain specificity of ZUFSP catalytic core (ZUFSP^310-578) compared to full-length Mug105. Both DUBs were tested against K48- and K63-linked Ub₂ for the indicated time points. e Pull-down analysis of ZUFSP (full-length and two N-terminal truncations) against a mixture of K63-linked Ub₄ and Ub₅ chains