Fig. 2 | Nature Communications

Fig. 2

From: A family of unconventional deubiquitinases with modular chain specificity determinants

Fig. 2

ZUFSP and Mug105 are ubiquitin-specific proteases with different chain specificity. a Activity assays with Ub-/UbL-AMC substrates shown as released fluorescence (RFU) over time (min) with ZUFSP or Mug105. Shown RFU values are the means of triplicates. b Fluorescent scan of a suicide probe reaction of ZUFSP or Ufsp2 with Cy5-Ub-PA (arrow) and Rho-Ufm1-PA (arrowhead). Asterisks (*) mark the shifted band after reaction. c Suicide probe reaction of ZUFSP or ATG4B with Ub-PA and LC3B-PA. Asterisks (*) mark the shifted band after reaction. d, e Linkage specificity analysis with ZUFSP. A panel of di-ubiquitin (d) or tetra-ubiquitin (e) chains was treated with ZUFSP for the indicated time points. f Time course of cleavage of K63-linked Ub6+ chains by full-length ZUFSP. g Linkage specificity analysis with Mug105. A panel of Ub2 was treated with Mug105 for the indicated time points. h Suicide probe reaction with Mug105 and K48-diUb-VME, K63-diUb-VME or Ub-PA. Arrows mark the shifted bands after reaction

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