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. Author manuscript; available in PMC: 2014 Aug 4.
Published in final edited form as: Cell Rep. 2012 Mar 19;1(4):309–316. doi: 10.1016/j.celrep.2012.02.006

Figure 3. NAE (UBA3 A171T) is less sensitive to MLN4924 and activates NEDD8 more efficiently in vitro.

Figure 3

MLN4924 titrations (2-fold dilutions) in NAE ATP:PPi exchange assay using 100 µM ATP (A) or 1 mM ATP (B). Kinetic measurements of ATP:PPi exchange by NAE and NAE (UBA3 A171T) in ATP (C) and PPi (D) titrations. ATP titrations used 1 mM PPi with the indicated concentrations of ATP. PPi titrations used 2 mM ATP with the indicated concentrations of PPi. All reactions included 50 cpm/pmol of [32P] PPi, 100 µM NEDD8, and 10 nM of NAE or NAE (A171T), and were incubated at 37°C for 30 minutes. See Figure S3 for raw data.