In retinal rods, photoexcited rhodopsin (R*) is inactivated upon phosphorylation by rhodopsin kinase and the subsequent binding of arrestin. We have studied the structural role of a cationic region of bovine arrestin (Val170-Arg182) using anti-peptide IgGs specifically recognizing this segment and the corresponding oligopeptide. Our results clearly indicate that amino acids Val170-Arg182 are shielded within the arrestin-rhodopsin-complex and very likely belong to a binding domain of arrestin for phosphorylated R*. The purified anti-peptide IgGs strongly reacted with isolated arrestin but did not recognize arrestin when bound to phosphorylated R*. In agreement with these experiments, the oligopeptide Val170-Arg182 was found to compete with arrestin for binding to phosphorylated R*. Increasing concentrations of this peptide caused an oligomerization of phosphorylated rhodopsin when illuminated by white light as well as in the dark. Unphosphorylated rhodopsin did not oligomerize up to a 400-fold molar ratio of peptide/rhodopsin. Limited proteolysis of the phosphorylated carboxy-terminus of rhodopsin with endoproteinase Asp-N caused a significant decrease in the peptide-induced formation of oligomers. Therefore, Val170-Arg182 of bovine arrestin probably interacts with the phosphorylated carboxy-terminus of rhodopsin. The data presented support the proposal of Palczewski et al. (1991c) considering the region Lys163-Arg182 in bovine arrestin to be a possible binding domain for phosphorylated R*.