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EC50 for melibiose displacement of D2G binding. With SLM-8100/DMX fluorescence spectrophotometer, displacements of D2G by melibiose at increasing concentrations were measured on a time trace with λEx 290 nm and λEm 500 nm and expressed as percentages of a total bound D2G. EC50 in the absence (square) or presence of Na+ (circle) or Li+ (triangle) was determined by hyperbolic fitting, and appKd for melibiose was calculated by the equation: appKd = EC50/{1 + [D2G]/Kd for D2G} (Table 1).
