Cloning of a melatonin-related receptor from human pituitary

• Published in: FEBS letters Vol. 386; no. 2; pp. 219 - 224
• Main Authors: Reppert, Steven M., Weaver, David R., Ebisawa, Takashi, Mahle, Cathy D., Kolakowski, Lee F.
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 20.05.1996
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Biological Evolution; Cell Line, Transformed; Cercopithecus aethiops; Cloning, Molecular; G protein-coupled receptor; GTP-Binding Proteins - metabolism; Human pituitary; Humans; Ligands; Melatonin; Molecular Sequence Data; Nerve Tissue Proteins - chemistry; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; Pituitary Gland - metabolism; Protein Conformation; Rats; Receptors, Cell Surface - chemistry; Receptors, Cell Surface - genetics; Receptors, Cell Surface - metabolism; Receptors, G-Protein-Coupled; Receptors, Melatonin; RNA, Messenger; G protein-coupled receptor; Melatonin; Human pituitary
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(96)00437-1

We have cloned an orphan G protein-coupled receptor from a human pituitary cDNA library using a probe generated by PCR. The cDNA, designated H9, encodes a protein of 613 amino acids that is 45% identical at the amino acid level to the recently cloned human Mel 1a and Mel 1b melatonin receptors. Structural analyses of the encoded protein and its gene, along with phylogenetic analysis, further show that H9 is closely related to the G protein-coupled melatonin receptor family. Unusual features of the protein encoded by H9 include a lack of N-linked glycosylation sites and a carboxyl tail >300 amino acids long. H9 transiently expressed in COS-1 cells did not bind [ 125I]melatonin or [ 3H]melatonin. H9 mRNA is expressed in hypothalamus and pituitary, suggesting that the encoded receptor and its natural ligand are involved in neuroendocrine function.