Human plasma afamin, the fourth member of the albumin gene family, is shown to be a specific binding protein for vitamin E. A radio ligand-binding assay followed by Scatchard and Hill analysis are used to show that afamin has a binding affinity for both alpha-tocopherol and gamma-tocopherol, two of the most important forms of vitamin E, in vitro. The binding-dissociation constant was determined to be 18 microM, indicating that afamin plays a role as vitamin E carrier in body fluids such as human plasma and follicular fluid under physiological conditions. Additionally, we demonstrate that afamin has multiple binding sites for both alpha- and gamma-tocopherol. Due to the large binding capacity of afamin for vitamin E, it might take over the role of vitamin E transport in body fluids under conditions where the lipoprotein system is not sufficient for vitamin E transport. To confirm the experimental results, we performed homology modeling and docking calculations on the predicted tertiary structure, which showed coincidence between calculated and in vitro results.