When hCG was receptor-bound, only 2 epitopes (i.e. beta 3 and beta 5) out of its previously mapped total of 14 surface epitopes (10, 11, 12) could be detected by the respective 125I-MCA. Clearly, this indicates a non-random orientation of hCG in this particular state (1). Now we report that on receptor-bound desialylated (asialo-hCG) as well as on receptor-bound deglycosylated hCG (degly-hCG), the beta 3 and beta 5 epitopes were inaccessible for 125I-MCA as were the remaining epitopes, although both variants, when not receptor-bound, were indistinguishable from native hCG with respect to number and topography of epitopes. Thus, the carbohydrate (CHO) units of hCG neither seem to be part of these 14 antigenic sites nor to contribute to the affinity of receptor binding: both variants had even higher affinities than native hCG. However, since the CHO are known to be obligatory for triggering the postreceptor responses of hCG-stimulated target cells, as seen by the 50% reduced or totally abolished biological potency of asialo-hCG and degly-hCG, respectively, the here demonstrated clear-cut differences in epitope accessibility can be related to differences in receptor-bound orientations which reflect signal transduction-competent and incompetent modes of interaction with the receptor. We believe that the CHO, while not contributing to receptor binding per se, function to assure correct positioning of hCG in the ligand recognition domain to allow for proper protein-protein interactions between ligand and receptor and thus optimal activation and hence transduction of the external signal to the cell's interior. In addition, the fact that most of the surface epitopes were masked on receptor-bound hCG, represents the first experimental support for the sofar unproven hypothesis that this unusually long (i.e. 341 amino acid residues) extracellular N' terminal domain of the recently cloned hCG receptor (hCG-R) (23,24), is indeed involved in ligand recognition.