Supplementary Figure 3: SPR analyses of MDM2 and MDMX variants binding affinities for UbcH5B–Ub
From: Structural analysis of MDM2 RING separates degradation from regulation of p53 transcription activity
(a) Representative sensorgrams (left) and binding curves (right) for GST-MDM2 398-C variants with UbcH5B–Ub in the presence of UbΔGG are shown. Only sensorgram is shown for GST-MDM2 398-C variants that displayed no measurable UbcH5B–Ub binding in the presence of UbΔGG. (b) Representative sensorgrams (left) and binding curves (right) for GST-MDMXR variants with UbcH5B–Ub are shown. Wild type MDMX displayed no UbcH5B–Ub binding up to 100 μM UbcH5B–Ub whereas both K478R and N448C, K478R mutants exhibited UbcH5B–Ub binding. However, Kd could not be estimated due to the weak binding affinity. All experiments in a and b were performed in duplicates.
