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Activation of NEDD8 and SUMO-2 by their cognate E1 enzymes. 15N-enriched NEDD8 (A) or SUMO-2 (B) were incubated with their respective E1 enzymes (APPBP1/UBA3 for NEDD8 and SAE1/SAE2 for SUMO-2) in the presence of MgCl2, ATP, and MESNa. The reactions were aliquoted and stopped at different time points by adding EDTA. An internal (unlabeled) standard was added, and the samples were subjected to ESI-MS. The representative spectral regions are shown.
