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Nedd4-2 forms thioester-linked polyubiquitin chains. Left, autoradiogram of 1-min reactions containing 40 nm each of Uba1, Ubc5B, and GST-Nedd4-2 or GST-Nedd4-2HECT resolved under non-reducing conditions, as indicated under “Materials and methods.” Right, parallel 1-min reactions resolved under reducing conditions. The positions of mobility standards are shown to the left; the position for free 125I-ubiquitin is shown to the right.
