Fig. 4: Ubiquitination of authentic substrate protein via the SUE1 strategy.
From: Structure-guided engineering enables E3 ligase-free and versatile protein ubiquitination via UBE2E1
a Top, SDS‒PAGE analysis of SUE1-mediated monoubiquitination of α-Synuclein (α-Syn) at K43 using In-gel fluorescence for clear visualization of ubiquitination. The model of α-Synuclein was derived from the NMR structure of the wild-type α-Synuclein (PDB: 1XQ8). Bottom, deconvoluted ESI-MS of the purified monoubiquitinated α-Synuclein (α-Syn-Ub). b Top, SDS‒PAGE analysis of SUE1-Mediated K63-linked ubiquitin chain modification of p53 at K24 and asterisk (*) denotes an impurity in the p53-stock. The p53 model was derived from the predicted structure of wild-type p53 in the AlphaFold Protein Structure Database (AF-P04637-F1-model_v4). Bottom, deconvoluted ESI-MS of the purified diubiquitinated p53 (p53-diUb). c Top, SDS‒PAGE analysis of SUE1-mediated monoubiquitination of p53 at K386 and asterisk (*) denotes an impurity in the p53-stock. Deconvoluted ESI-MS of the purified ubiquitinated products. Bottom, deconvoluted ESI-MS of the purified monoubiquitinated p53 (p53-Ub). Gel images shown in (a–c) are representative of independent biological replicates (n = 2). Source data are provided as a Source Data file.
