Molecular cloning, sequence and functional expression of the cDNA for the human thyrotropin receptor

• Published in: Biochemical and biophysical research communications Vol. 165; no. 3; pp. 1184 - 1190
• Main Authors: Nagayama, Yuji, Kaufman, Keith D., Seto, Pui, Rapoport, Basil
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 29.12.1989; Elsevier
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; Cell Line; Cloning, Molecular; Codon; Cricetinae; Cyclic AMP - metabolism; DNA - genetics; Fundamental and applied biological sciences. Psychology; Gene Expression; genes; Genes. Genome; Glycosylation; Humans; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Molecular Weight; Receptors, LH - genetics; Receptors, Thyrotropin - genetics; Sequence Homology, Nucleic Acid; thyroid; Transfection; TSH; PCR; CHO; Human; Cell culture; Nucleotide sequence; Rodentia; Gene expression; Glycoprotein hormone; CHO cell line; Vertebrata; Mammalia; Complementary DNA; Adenohypophyseal hormone; Radioimmunoassay; Molecular cloning; Nucleic acid; Hamster; Aminoacid sequence; Hormonal receptor
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/0006-291X(89)92727-7

We screened a human thyroid cDNA library with two synthetic oligonucleotides based on the reported amino acid sequence of the 3rd and 4th transmembrane domains of a putative human TSH receptor and related receptors. The nucleotide sequence of a 4 kb clone revealed an open reading frame of 764 amino acids (86,816 Daltons) with a putative signal peptide, seven transmembrane domains, five potential glycosylation sites, and a very short intracytoplasmic region. Homology with the extracellular domain of the pig LH/CG receptor was only 33%. Chinese hamster ovary cells stably transfected with this cDNA in an expression vector generated a functional receptor, able to activate adenylate cyclase, specifically in response to TSH stimulation.