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. Author manuscript; available in PMC: 2017 Feb 18.
Published in final edited form as: Nature. 2016 Aug 18;536(7616):304–308. doi: 10.1038/nature19071

Extended Data Figure 2. Comparing Strategies for Crosslinking the E2-SUMO Thioester Mimic and Substrate PCNA.

Extended Data Figure 2

a, Chemical structures of the proposed tetrahedral intermediate formed during PCNA Lys164 attack of E2Ubc9-SUMO thioester (left), a BMOE cross-link (middle) or an EDT cross-link (right) between E2Ubc9-SUMO C93 and PCNA K164C. Indicated distances were estimated with ChemDraw15 (PerkinElmer). b, Control non-reducing SDS-PAGE panel for Fig. 1a showing mock treated PCNA K127R/K164C (DMSO instead of EDT in DMSO) is unable to accept transthioesterification of SUMO at position 164. c, SDS-PAGE analysis of the 5 ml fractions from the final preparative Superdex200 gel-filtration purification of the E2Ubc9-SUMO-EDT-PCNA/ E3Siz1(167–449)-SUMO complex. For gel source data, see Supplementary Figure 1.