The cellular transport, metabolism and biological activity of retinoids are mediated by their specific binding proteins and nuclear receptors. For an understanding of the mode of action of retinoids with potential cancer chemopreventive or other biological activity, it is important to study their interactions with these binding proteins and receptors. In our attempts to understand the action of N-(4-hydroxyphenyl)retinamide (4HPR) and other retinamides in the prevention of cancer, we observed that 4HPR binds to a serum protein with a molecular size of approximately 20,000. The retinoid, however, did not show any binding affinity for cellular retinol-binding protein (CRABP) or for cellular retinoic acid-binding protein (CRABP). However, it showed binding affinity for the nuclear receptors of retinoic acid (RARs) equivalent to 15% of that of retinoic acid. The physicochemical properties of the 4HPR binding protein in the serum were identical to those of serum retinol binding protein (RBP). Antibodies against RBP quantitatively immunoprecipitated the protein-4HPR complex, confirming that the retinoid specifically binds to RBP. Although retinol and 4HPR cross-competed for RBP binding, N-phenylretinamide, in which the 4-hydroxyl group is absent, and N-(4-methoxyphenyl)retinamide, a major cellular metabolite of 4HPR, in which the hydroxyl group is blocked, did not show affinity for the binding protein. The results indicate that the hydroxyl group of 4HPR is essential for binding of this type of retinoid to RBP. Thus, our studies suggest that serum transport of 4HPR may be facilitated by RBP. To bind more efficiently to CRBP, CRABP, or RARs/RXRs, the retinoid may require further metabolic change.