Identification of Mel(1a) melatonin receptors in the human embryonic kidney cell line HEK293: Evidence of G protein-coupled melatonin receptors which do not mediate the inhibition of stimulated cyclic AMP levels

Binding assays using 2-[I-125]iodomelatonin revealed high-affinity, guanosine 5'-O-(3-thiotriphosphate) sensitive, melatonin binding sites (B-max 1.1 fmol/mg protein) in the human embryonic kidney cell line HEK293. Competition studies using the selective melatonin receptor antagonist luzindole and RT-PCR techniques identified these sites as human Mel(1a), melatonin receptors, Challenge of HEK293 cells with 1 mu M melatonin had no effect on forskolin stimulated cyclic AMP levels, whereas in HEK293 cells engineered to stably over-express the human Mel(1a) melatonin receptor (B-max > 400 fmol/mg protein) melatonin dose-dependently inhibited stimulated cyclic AMP levels (IC50 7.7 pM. These data may indicate that certain tissues, expressing low levels of G protein-coupled melatonin receptors, do not display melatonin mediated inhibition of cAMP. (C) 1997 Federation of European Biochemical Societies.