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Binding of 11-cis retinol to X2IRBP of wild-type sequence. The titrations monitored the enhancement of retinol fluorescence (excitation 330 nm, emission 480 nm) and quenching of endogenous protein fluorescence (excitation 280 nm, emission 340 nm). Enhancement: N = 0.95 ± 0.03 and Kd = 0.077 ± 0.015 μM. Quenching: N = 0.93 ± 0.02 and Kd = 0.042 ± 0.009. The calculated number of binding sites and dissociation constants are summarized in Table 2.
