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. 1998 Aug 18;95(17):9802–9807. doi: 10.1073/pnas.95.17.9802

Figure 5.

Figure 5

Fluorescence emission spectra of MIANS-labeled permease mutants and effect of TDG. Each single-Cys permease mutant was labeled with MIANS while bound to avidin-Sepharose. Reactions were terminated with DTT, and the excess reagent was removed by extensive washing before elution of the labeled protein from the affinity column. Emission spectra of MIANS-labeled permease V331C (A), E126A/V331C (B), R144A/V331C (C), and R144K/V331C (D) [40 μg of pure protein in 0.5 ml of 50 mM KPi (pH 7.5)/150 mM NaCl/0.02% DM] were measured (excitation 330 nm) at 22°C after preincubation with a given concentration of ligand as follows: curve 1, 10 mM sucrose; curve 2, 10 mM TDG; curve 3, 0.2 mM TDG.