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Effect of a membrane-localized quencher DPA on TMR-labeled KcsA mutants at pH 4 and pH 7. A, shown are emission spectra for A132C in the presence and absence of DPA. The data were normalized at 570 nm in the absence of DPA. B, fluorescence intensities in the presence and absence of DPA at pH 4 and pH 7 at 570 nm are shown. The histogram shows results normalized for protein concentration (mean ± S.D., n = 3–6). *, p < 0.05 compared with quenching ratio by DPA at pH 7.
