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Manganese ion dependency of enzyme activity of α551 and α551.D286A. GST-Rac (1 μg) was incubated with α551 (1 nM; B) or α551.D286A (500 nM; A) in the presence of UDP–[14C]N-acetylglucosamine (10 μM) with the indicated concentrations of MnCl2 for 30 min. Labeled proteins were then analyzed by SDS-PAGE and phosphorimaging (shown).
