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. 2010 Aug 3;285(43):33457–33462. doi: 10.1074/jbc.M110.135582

FIGURE 1.

FIGURE 1.

Effect of Ubc9 and Ubc9 mutants that have reduced affinity for E1 on formation of the Ubc9∼SUMO thioester. A, the E1-catalyzed formation of the Ubc9∼SUMO thioester conjugate in the presence of wild-type and mutant Ubc9 at the five indicated concentrations of wild-type and mutant Ubc9 and the three indicated incubation times. B, quantification of the assay results in A. C, formation of RanGAP1-SUMO isopeptide complex in the presence of wild-type and mutant Ubc9. The conjugation assays were carried out at the five indicated concentrations of wild-type and mutant Ubc9 and the three indicated incubation times. All other conditions of the reactions are identical among all reactions in A and C. Details of the assay conditions are as described under “Experimental Procedures.” Loading control gels for wild-type and mutant Ubc9 proteins are shown as the bottom gels of A and C. D, quantification of the assay results in C.