Fig. 4: The effect of Ub4a binding on the in vitro K48-linked enzymatic chain assembly. | Nature Communications

Fig. 4: The effect of Ub4a binding on the in vitro K48-linked enzymatic chain assembly.

From: Mechanism of selective recognition of Lys48-linked polyubiquitin by macrocyclic peptide inhibitors of proteasomal degradation

Fig. 4

SDS PAGE gels of K48-linked Ub chain assembly by E2-25K enzyme (GST-fused), and their quantitation. Shown are the results for select time points in the reaction (a) in the presence of equimolar (to Ub) amount of Ub4a and (b) without Ub4a, as control. Molecular weight (MW) values are shown in kDa. The normalized intensities of the Ub1-Ub4 bands are plotted in c and d respectively. The data for Ub1, Ub2, Ub3, and Ub4 are colored black, red, blue, and green, respectively. Shown are the mean values obtained in three attempts (n = 3) to quantitate gel bands using ImageJ; the error bars, centered on the mean values, represent standard deviations among these measurements. The gray dots depict the normalized intensities obtained in each of these attempts. The cartoon on the bottom illustrates the chain assembly reaction. Source data are provided as a Source Data file.

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