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. 2001 Mar 27;98(7):3710–3715. doi: 10.1073/pnas.061455898

Figure 2.

Figure 2

Retinol binding to human apo-CRBP III. The absorption spectra of the retinol–CRBP I (A) and retinol–CRBP III (B) complexes were recorded after the addition of a slight molar excess of all-trans-retinol to recombinant rat apo-CRBP I and human apo-CRBP III (9 and 7 μM, respectively), followed by equilibration in the dark for a few minutes at 20°C. (C) Spectrofluorometric titration of human CRBP III with retinol. Intrinsic protein fluorescence (%) of apo-CRBP III (0.8 μM) is plotted as a function of retinol concentration. (Inset) Fitting of the experimentally determined fractional saturation to a theoretical curve corresponding to the binding of ≈0.66 mol of ligand per mol of CRBP III, with an apparent Kd of 60 nM.