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Retinol binding to Xenopus IRBPs consisting of two contiguous modules. Binding of double modules IRBPs with all-trans retinol as followed during titrations by monitoring the increase in retinol fluorescence (excitation, 330 nm; emission, 480 nm). The concentration of the IRBP double module was: modules 1&2, 0.67 μM; modules 3&4, 0.57 μM. A) Xen IRBP modules 1&2 showed N = 2.45 ± 0.11 with Kd = 0.049 ± 0.023 μM. B) Xen IRBP modules 3&4 showed N = 1.43 ± 0.21 with Kd = 0.19 ± 0.05 μM.
