Interactions of retinol with binding proteins: studies with rat cellular retinol-binding protein and with rat retinol-binding protein.
The interactions of retinol with rat cellular retinol-binding protein (CRBP) and with rat serum retinol-binding protein (RBP) were studied. The equilibrium dissociation constants of the two retinol-protein complexes (Kd) were found to be 13 x 10(-9) and 20 x 10(-9) M for CRBP and for RBP, respectively. The kinetic parameters governing the interactions of retinol with the two binding proteins were also studied. It was found that although the equilibrium dissociation constants of the two retinol-protein complexes were similar, retinol interacted with CRBP 3-5-fold faster than with RBP; the rate constants for dissociation of retinol from CRBP and from RBP (koff) were 0.57 and 0.18 min-1, respectively. The rate constants for association of retinol with the two proteins (kon) were calculated from the expression: Kd = koff/kon. The kon's for retinol associating with CRBP and with RBP were found to be 4.4 x 10(7) and 0.9 x 10(7) M-1 min-1, respectively. The data suggest that the initial events of uptake of retinol by cells are not rate-limiting for this process and that the rate of uptake is probably determined by the rate of metabolism of this ligand. The data indicate further that the distribution of retinol between RBP in blood and CRBP in cytosol is at equilibrium and that intracellular levels of retinol are regulated by the levels of CRBP.
Noy
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Department of Medicine, Cornell University Medical College, New York, New York 10021.
Blaner
W S
WS
D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D007700:Kinetics; D008081:Liposomes; D008099:Liver / Q000378:metabolism*; D008433:Mathematics; D010713:Phosphatidylcholines; D011485:Protein Binding; D051381:Rats; D012177:Retinol-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D054840:Retinol-Binding Proteins, Cellular; D013050:Spectrometry, Fluorescence; D014801:Vitamin A / Q000378:metabolism*
D008081:Liposomes; D010713:Phosphatidylcholines; D012177:Retinol-Binding Proteins; D054840:Retinol-Binding Proteins, Cellular; D014801:Vitamin A; C017251:1,2-oleoylphosphatidylcholine