Major histocompatibility complex (MHC)-restricted recognition of antigen by T lymphocytes involves the formation of a complex composed of the T cell receptor, antigen, and restricting MHC molecule. To elucidate the interactions occurring within the antigen recognition complex, we have evaluated the ability of a panel of cell lines expressing mutated I-Ak molecules to function in the recognition by T hybridoma cells of two distinct peptide antigens. Our results indicate that while alterations along the entire length of the proposed helical structure in the carboxyterminal half of the beta 1 domain interfere with the I-Ak-restricted recognition of human fibrinopeptide B, mutations which affect recognition of hen egg lysozyme/I-Ak fall almost exclusively in the central portion of the helix. On the basis of these and previous results, we propose a "T cell receptor-mediated peptide exchange model" for formation of the antigen recognition complex.