FIGURE 9.
Phosphorylation of WT CopA with Pi and subsequent decay. A, phosphorylation was started by the addition of 50 μm [32P]Pi to WT CopA in conjunction with 1 mm BCS (•) or 40 μm CuCl2 (○), or following a 2-min preincubation with 40 μm CuCl2 (▾). B, Pi concentration dependence of the phosphoenzyme equilibrium level, in the presence of 1 mm BCS. Incubation was for 5 min at 60 °C. C, decay was started with a 1 mm non-radioactive Pi chase added to [32P]phosphoenzyme obtained by a 5-min incubation with 50 μm [32P]Pi in the absence of both BCS and CuCl2. The chase medium contained either 1 mm BCS (•) or 40 μm CuCl2 (○). The temperature was 60 °C.