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E1 and E2-25K-catalyzed polymerization of hydrophobic patch mutants of Ub and of K48-linked polyUbs. SDS-PAGE gels of in vitro E1 and E2-25K-catalyzed polymerization reactions for UbWT and its mutants (A-D) or preassembled polyUbs (E,F), as indicated. The reactions were performed and analyzed as detailed in Supporting Information. The running positions of the E2 and select Ub polymers are indicated by arrows.
