Extended Data Fig. 3: Design and purification of the stable intermediate structures.
From: Structural insights into Ubr1-mediated N-degron polyubiquitination
a, A schematic representation of the transition state of the initiation step. The side chain of a Lys residue on Degron attacks the thioester bond of Ubc2–Ub. The inset shows the designed intermediate structure mimicking the transition state of the initiation step. b, A schematic representation of the transition state of the elongation step. The side chain of Lys48 on Ub-Degron attacks the thioester bond of Ubc2–Ub. The inset shows the designed intermediate structure mimicking the transition state of the elongation step. c, A brief synthetic route of the intermediate structure mimicking the transition state of the elongation step. d, A gel filtration chromatogram of Ubr1 (left) and an SDS–PAGE gel of purified Ubr1 and designed stable intermediate structures Ubc2-Ub-Degron and Ubc2-Ub-Ub-Degron (right).
