AbstractMelatonin receptors (MT₁ and MT₂ in humans) are family A G protein–coupled receptors that respond to the neurohormone melatonin to regulate circadian rhythm and sleep. Numerous efforts have been made to develop drugs targeting melatonin receptors for the treatment of insomnia, circadian rhythm disorder, and cancer. However, designing subtype-selective melatonergic drugs remains challenging. Here, we report the cryo-EM structures of the MT₁–G_i signaling complex with 2-iodomelatonin and ramelteon and the MT₂–G_i signaling complex with ramelteon. These structures, together with the reported functional data, reveal that although MT₁ and MT₂ possess highly similar orthosteric ligand-binding pockets, they also display distinctive features that could be targeted to design subtype-selective drugs. The unique structural motifs in MT₁ and MT₂ mediate structural rearrangements with a particularly wide opening on the cytoplasmic side. G_i is engaged in the receptor core shared by MT₁ and MT₂ and presents a conformation deviating from those in other G_i complexes. Together, our results provide new clues for designing melatonergic drugs and further insights into understanding the G protein coupling mechanism.