Cloning and Functional Expression of a Vascular Smooth Muscle Endothelin 1 Receptor

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 88; no. 8; pp. 3185 - 3189
• Main Authors: Lin, Herbert Y., Kaji, Eugene H., Winkel, Glen K., Ives, Harlan E., Lodish, Harvey F.
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 15.04.1991; National Acad Sciences
• Subjects: A10 cells; Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; Blotting, Northern; Calcium - physiology; Cell lines; Cell receptors; Cell structures and functions; Cells; CHO cells; Cloning, Molecular; Complementary DNA; COS cells; Cultured cells; DNA - genetics; endothelin; Endothelins; Endothelins - metabolism; Fundamental and applied biological sciences. Psychology; Gene Expression; GTP-Binding Proteins - physiology; Hormone receptors. Growth factor receptors. Cytokine receptors. Prostaglandin receptors; Molecular and cellular biology; Molecular Sequence Data; Muscle, Smooth, Vascular - physiology; Rats; Receptors; Receptors, Cell Surface - genetics; Receptors, Cell Surface - physiology; Receptors, Endothelin; RNA; Signal Transduction; Smooth muscle myocytes; Endothelin; Rat; Nucleotide sequence; Rodentia; Peptide hormone; Smooth muscle; Gene expression; Vertebrata; Membrane receptor; Mammalia; Cell line; Vasoconstrictor agent; Molecular cloning; Aminoacid sequence; Hormonal receptor
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.88.8.3185

By screening a cDNA library derived from the A10 rat vascular smooth muscle cell line for functional expression in COS cells, we have isolated a high-affinity receptor for endothelin 1 (Kd= 476 pM) and endothelin 2. The affinity of the cloned endothelin receptor for endothelin 3 is >100 times less in A10 cells and in a CHO cell line stably transformed by the endothelin receptor cDNA. The 426-amino acid receptor polypeptide has seven putative hydrophobic transmembrane domains and is presumed to be a member of the family of guanine nucleotide-binding regulatory (G) protein-coupled receptors. Microinjection of in vitro transcripts of the cloned cDNA into CHO cells confers a transient increase in intracellular calcium in response to endothelin 1, indicating that the receptor is functional and couples to the appropriate G protein(s). RNA analysis reveals high expression in rat lung and heart, tissues known to exhibit binding to iodinated endothelin 1.