Identification of the MINDY Family of DUBs
(A) Schematic representation of the domain structure of human FAM63A.
(B) Halo-tagged FAM63A (388–426) coupled to HaloLink resin was incubated with tetraUb of the indicated linkage types. The captured materials were separated on 4%–12% SDS-PAGE gel and silver stained.
(C–H) DUB assays testing activity and specificity of polyUb cleavage by FAM63A (C), putative catalytic domain of FAM63A (110–384) (D) (asterisk indicates FAM63A), full-length FAM63B (E), YPL191C/MIY1 (F), YGL082W (G), and full-length human FAM188A (H); 1.6 μM of DUBs were incubated with 2.2 μM of tetraUb chains for the indicated time.
(I) Phylogenetic tree of MINDY family DUBs based on alignment of catalytic domains (Figure S2B).
See also Figure S1.