We have synthesized peptides corresponding to the known sequence of the rat dopamine D1 receptor and measured the effects on binding of the Gs protein to the activated receptor. Peptides corresponding to the second and third cytoplasmic loop (amino acids 160-180 and 260-276), as well as the N-terminal part of the carboxy terminus (amino acids 375-391) were effective in preventing Gs binding to the activated receptor. Peptides representing other surface regions, e.g., the first cytoplasmic loop (87-100), the C-terminus (400-420 and 440-460) and all extracellular surface peptides (6-22, 120-137, 210-224 and 338-352) are not involved in rat dopamine D1 receptor-Gs protein interaction.