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. Author manuscript; available in PMC: 2018 Apr 20.
Published in final edited form as: Cell Chem Biol. 2017 Mar 16;24(4):443–457.e6. doi: 10.1016/j.chembiol.2017.02.013

Figure 2.

Figure 2

Ub2PT crosslink to polyUb-binding proteins. (A) K48-linked or K63-linked diUbPT was crosslinked to Dsk2, Rad23, Rpn10, or a control protein Rpn12 according to protocol described in Figure 1B. (B) Model of Rad23-UBA2 molecular structure (cyan) in complex with K48-Ub2, distal Ub (green) and proximal Ub (orange) based on PDB: 1ZO6. In this orientation, proximity of Leu73 and of Leu8 (magenta sticks) of the distal Ub to the receptor are apparent. (C) RAP80-tUIM selectively crosslinks to K63-Ub2PT(8). In this case, RAP80-tUIM efficiently crosslinked to K63-Ub2PT(8) but not to a dimer linked via Lys48, demonstrating that UbPT allows for trapping of K63-linkage selectivity. (D) Crystal structure of RAP80-tUIM (cyan) in complex with K63-Ub2 (distal Ub green and proximal Ub orange) from PDB:3A1Q, highlights that Leu8 (magenta stick) on the distal Ub is in close proximity to the ligand.