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. 2019 Aug 5;9:11338. doi: 10.1038/s41598-019-47824-y

Figure 3.

Figure 3

The C154G mutation improves the stability of the inverted LacY in DDM micelles. (A) Representative melting curves using the CPM dye binding assay for WT and C154G LacY isolated from PE-lacking cells. LacY was diluted to 26 μg/mL in 50 mM Tris-HCl (pH 7.5), 100 mM NaCl containing 0.05% DDM. (B) Intrinsic Trp fluorescence intensity and (C) changes in the maximum emission wavelength (λmax) of the fluorescence emission spectrum as a function of the temperature for WT and C154G LacY isolated from PE-lacking cells. Protein were diluted to 10 μM in 50 mM Tris-HCl (pH 7.5), 100 mM NaCl containing 0.05% DDM. Solid lines represent log(agonist) vs. response (Variable slope) fits of the data. In all cases the experiments were repeated three times and the data represent mean values ± SD.