title: Interactions of retinol with binding proteins: implications for the mechanism of uptake by cells.
abstract: The kinetic parameters of the interaction of retinol with retinol binding protein (RBP) were studied. The rate constant for association of retinol with the protein (ka) was found to be 1.5 X 10(6) M-1 min-1. The rate constant for dissociation (kd) from the protein was determined by studying the transfer of retinol from RBP to lipid bilayers. It was found that such transfer proceeds via the aqueous phase and its rate-limiting step is the dissociation of retinol from the binding protein. The rate of transfer therefore represents the rate of dissociation. The kd was 0.112 min-1. These values were validated further by the following consideration. The equilibrium dissociation constant of RBP and retinol can be calculated from the expression Kd = kd/ka. The calculated value was 7.5 X 10(-8) M. Kd was also measured directly by fluorometric titration and was found to be 7 X 10(-8) M. The relative avidities of retinol for RBP, the complex RBP-transthyretin (RBP-TTR), and serum albumin were also studied. It was found that binding of RBP to TTR increased its avidity for retinol by about 2-fold. The avidity of albumin for retinol was 30-fold lower than that of RBP. The data imply that retinol spontaneously and rapidly dissociates from sites on binding proteins, which indicates that the vitamin can freely move in vivo between physiologic compartments with avidities for it.
authors.affiliation: Department of Medicine, Cornell University Medical College, New York, New York 10021.
authors.forename: N
authors.initials: N
authors.lastname: Noy
authors.forename: Z J
authors.initials: ZJ
authors.lastname: Xu
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D011228:Prealbumin / Q000378:metabolism; D051381:Rats; D012177:Retinol-Binding Proteins / Q000378:metabolism*; D014801:Vitamin A / Q000378:metabolism*
chemical_list: D011228:Prealbumin; D012177:Retinol-Binding Proteins; D014801:Vitamin A