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Fig. 2 | Cell Division

Fig. 2

From: COP9-Signalosome deneddylase activity is enhanced by simultaneous neddylation: insights into the regulation of an enzymatic protein complex

Fig. 2

ATP-depletion resulted in the inhibition of CSN deneddylase activity. a 10 nM His-6-Cul1/ROC1 were pre-neddylated for 30 min at 30°C. Then 10 μg HeLa cells extract were added at 30°C (time plot: lanes 36); or alternatively, ATP-trap mix was added for 45 min at 30°C, followed by the addition of 10 μg HeLa cells extract at 30°C (time plot: lanes 811). b 3 nM His-6-Cul1/ROC1 was pre-neddylated for 30 min at 30°C. Then 100 nM purified CSN was added at 20°C (time plot: lanes 36); or alternatively, ATP-trap mix was added for 45 min at 30°C, followed by the addition of 100 nM purified CSN at 20°C (time plot: lanes 811). c 10 nM His-6-Cul1/ROC1 was incubated as described in b; time plots were for 1.5–7.5 min, and not for 5–40 min as in b. Blotting was directed against NEDD8, and not against Cul1, as described elsewhere.

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