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pH dependence of Δ-IR intensity change at 1,742 cm−1 measured with LacYww in the absence or presence of NPG (filled red circles) or E325A LacY (open red circles), and Kd values for NPG binding to WT LacY (filled green circles), E325A LacY (open green circles), or LacYww (filled cyan circles). Kd values were calculated as the ratio of rate constants (koff/kon) measured by stopped flow (12, 35).
