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. Author manuscript; available in PMC: 2011 May 4.
Published in final edited form as: Biochemistry. 2010 May 4;49(17):3658–3666. doi: 10.1021/bi100026a

Figure 7.

Figure 7

Transfer of the metalloid from ArsD occurs during ArsA catalysis. As(III) transfer assays were performed as described in Materials and Methods. MBP-ArsD109 was bound to an amylose resin, and then the column was charged with sodium arsenite to saturate ArsD109. Next purified ArsA or BSA was applied to the columns with the indicated nucleotides (5 mM) at room temperature unless otherwise noted. (A) Ten fractions were collected and analyzed by SDS–PAGE. Nearly all of the BSA or ArsA eluted in fraction 2, and fraction 9 contained nearly all of theMBP-ArsD109. Nucleotides: (B) ArsA with MgATP,(C) BSA with MgATP,(D) ArsA with MgADP, (E)ArsA with MgATP-γ-S, and(F)ArsA with MgATP at 4 °C. The concentration of protein (open bars) and As(III) (●) was quantified in each fraction. The error bars represent the standard deviation from three assays.