Fig. 2: The UBS independent role of the linker connecting the UBS to UIS.

a SDS–PAGE analysis showing the effect of UBA5 truncations or deletion on the transfer of UFM1 to UFC1. The gel is representative of at least two independent experiments. b ITC experiment of UFC1 binding to UBA5 Δ349−389. The top graph represents raw data of heat flow versus time. The area under the peaks of the upper panel was integrated and plotted as kcal per mole of UFC1 as a function of binding stoichiometry in the bottom panel. Thermodynamic parameters are summarized in Supplementary Table 2. c SDS–PAGE analysis showing the discharge of UBA5 WT or (1–347) by free cysteine (100 mM). The gel is representative of at least two independent experiments. d SDS-PAGE analysis showing the effect of UBA5(347–404) on the transfer of UFM1 to UFC1 from UBA5(1–347). e Three independent experiments as shown in d were performed to quantify the effect of adding the UBA5 fragment in trans on UFM1 transfer to UFC1. Data are represented by mean ± SD.