Fig. 5
From: Reactive-site-centric chemoproteomics identifies a distinct class of deubiquitinase enzymes
ZUFSP interacts with ubiquitin and reacts with Ub ABPs. a Domain structure of ZUFSP. Domain prediction from SMART (http://smart.embl-heidelberg.de)44. Figure produced using Illustrator for Biological Sequences (http://ibs.biocuckoo.org)70. b A representative MS/MS spectrum showing a peptide-spectrum matched to ZUFSP Cys-360-labeled peptide sequence. c Pull-down of FLAG-ZUFSP by Ub, SUMO2, or UFM1-conjugated agarose. Lysates from FLAG-ZUFSP-overexpressing cells were enriched on the indicated resins and analyzed by anti-FLAG immunoblotting. d Reactivity of full-length ZUFSP toward ABPs. Purified ZUFSP (wt, C360A, or H491A mutant) was treated with the indicated ABP (Ub-VPS, Ub-VS, Ub-VME, Ub-PA, or SUMO2-VS [S2VS]), then analyzed by SDS-PAGE and stained with Coomassie Brilliant Blue. e Reactivity of the catalytic domain of ZUFSP toward ABPs. Purified ZUFSP(298–578) (wt or C360A mutant) was treated with ABP, then analyzed by SDS-PAGE, and stained with Coomassie Brilliant Blue
