MOLECULAR-CLONING AND FUNCTIONAL EXPRESSION OF A HUMAN THYROTROPIN-RELEASING-HORMONE (TRH) RECEPTOR GENE

In this study, we isolated genomic DNA fragments coding for the human thyrotropin-releasing hormone (TRH) receptor. Analysis of the nucleotide sequence revealed that the human TRH receptor gene had an exon-intron structure comprising at least two exons. A polypeptide encoded by the gene consisted of 398 amino acid residues with putative seven transmembrane domains. It showed high homology as a whole amino acid sequence with the rat and mouse TRH receptors except for considerable variation in the C-terminal region. Chromosomal mapping study indicated that the human TRH receptor gene was assigned to chromosome 8. Chinese hamster ovary (CHO) cells transfected with a DNA fragment containing the coding regions of the human TRH receptor bound with [H-3]TRH. This binding was inhibited by adding unlabeled TRH in a dose-dependent fashion. Scatchard analysis indicated that the transfected CHO cells expressed a single class of high affinity binding sites at a dissociation constant (K-d) of approximately 1 nM. These results demonstrated that the isolated gene encoded a specific TRH receptor with high affinity.