Extended Data Fig. 5: Preparation of covalent complexes of USP36 with Fubi and Ubiquitin probes for crystallography.
From: Molecular basis for ubiquitin/Fubi cross-reactivity in USP16 and USP36
a. Semi-synthesis of Ub-PA probe from Ub-MesNa (upper part) and characterization of these species by intact protein mass spectrometry (lower part). b. Intact mass spectrometry analysis of USP36 binding to Fubi- and Ub-PA probes, confirming 1:1 stoichiometry. c. Reactivity assessment of the shorter USP3681–424 construct with indicated probes as shown for the longer USP36 construct in Fig. 3c. The uncropped gel is provided as Source Data. d. Stability assessment of the shorter USP3681–424 construct complexed with Fubi or Ub probes as shown for the longer USP36 construct in Fig. 3f. Means and individual results of three samples are plotted. Tm, protein melting temperature.
