The interaction between retinol-binding proteins and prealbumins studied by fluorescence polarization.
The interaction between retinol-binding proteins and prealbumins of human and chicken was studied by fluorescence polarization techniques. The binding affinity between chicken plasma retinol-binding protein and chicken prealbumin was essentially the same as between the respective human proteins. Human urine retinol-binding protein displayed a similar affinity, though possibly slightly smaller than that of the human plasma protein, toward human prealbumin. Retinol-binding proteins and prealbumins of human and chicken have been found to cross-interact displaying an affinity similar to that displayed by the proteins of the same species. Solution of a binding equation which assumes identical, independent sites, indicated that the number of binding sites on prealbumin for retinol-binding protein is somewhat less than 2 with the human system, and in the neighborhood of 4 with the chicken system. A possible interpretation suggests that prealbumin possesses four identical binding sites for retinol-binding protein, one for each subunit, but that the binding is of a negative cooperative nature. A major share of the negative cooperativity is likely to result from steric hindrance induced by already bound retinol-binding protein molecules, which have a sizable volume compared to the volume of the prealbumin molecule. The cooperativity is likely to be more pronounced with the human system. Rotational relaxation times derived from Perrin plots suggest that 1:1 molecular complexes of retinol-binding proteins with prealbumins have a compact structure.
D000818:Animals; D001665:Binding Sites; D002645:Chickens; D006801:Humans; D007700:Kinetics; D046911:Macromolecular Substances; D008433:Mathematics; D011228:Prealbumin; D011485:Protein Binding; D012044:Regression Analysis; D012177:Retinol-Binding Proteins / Q000652:urine; D054839:Retinol-Binding Proteins, Plasma; D012709:Serum Albumin; D013045:Species Specificity; D013050:Spectrometry, Fluorescence; D013696:Temperature
D046911:Macromolecular Substances; D011228:Prealbumin; D012177:Retinol-Binding Proteins; D054839:Retinol-Binding Proteins, Plasma; D012709:Serum Albumin