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. 2009 May 5;284(28):18715–18725. doi: 10.1074/jbc.M109.015099

FIGURE 3.

FIGURE 3.

Na+ dependence of phosphorylation by ATP. Phosphorylation was carried out for 10 s at 0 °C in 20 mm Tris (pH 7.5), 3 mm MgCl2, 2 μm [γ-32P]ATP, 10 μm ouabain, 20 μg/ml oligomycin, and the indicated concentrations of Na+ added as NaCl with varying amounts of N-methyl-d-glucamine to maintain the ionic strength. Filled circle, wild type; crossed circle, ΔY; open triangle pointing upward, ΔYY; filled triangle pointing upward, ΔTYY; dotted triangle pointing upward, ΔWVEKETYY; dotted triangle pointing downward, YY-AA; dotted circle, Y1017F; open triangle pointing downward, Y1018F; dotted square, Y1017A; filled triangle pointing downward, Y1018A; open square, EKE-AAA; open diamond, R936A; crossed diamond, R935A; dotted diamond, GG-AA; filled square, K768M; crossed square, K768A. For comparison, the wild type is shown in all panels. Each line shows the best fit of the Hill equation, and the extracted K0.5(Na+) values are indicated in Table 1.