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. 2013 Jun 4;288(29):21320–21328. doi: 10.1074/jbc.M113.476242

FIGURE 4.

FIGURE 4.

Binding of AIPL1 and the FKBP-domain AIPL11–161 to farnesyl-Cys-AMCA probe. A, emission spectra of farnesyl-Cys-AMCA (0.17 μm) (1), AIPL1 (0.1 μm) (2), or AIPL1 (0.1 μm) mixed with farnesyl-Cys-AMCA (0.17 μm) (3) collected with excitation at 280 nm. Inset, Ni-NTA-purified AIPL1 stained with Coomassie Blue. B, FRET assay of [farnesyl-Cys-AMCA] binding to AIPL1 (λex 280 nm, λem 440 nm). Changes in farnesyl-Cys-AMCA fluorescence F − F0 (mean ± S.E. (error bars), n = 3) in the presence of increasing concentrations of AIPL1 were fit with an equation for binding with ligand depletion. C, emission spectra of AIPL11–161 (0.2 μm) alone (1) or mixed with farnesyl-Cys-AMCA (0.25 μm) (2) (λex 280 nm). Inset, Ni-NTA-purified AIPL11–161 and AIPL1162–328 stained with Coomassie Blue. D, changes in farnesyl-Cys-AMCA fluorescence F − F0 (mean ± S.E., n = 3) in the presence of increasing concentrations of AIPL11–161 fit with an equation for binding with ligand depletion.