Cloning and functional expression of a vascular smooth muscle endothelin 1 receptor.
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 greater than 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.
Lin
H Y
HY
Whitehead Institute for Biomedical Research, Cambridge, MA 02142.
Kaji
E H
EH
Winkel
G K
GK
Ives
H E
HE
Lodish
H F
HF
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002118:Calcium / Q000502:physiology; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D016232:Endothelins / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000502:physiology; D015870:Gene Expression; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000502:physiology*; D051381:Rats; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000502:physiology; D017466:Receptors, Endothelin; D015398:Signal Transduction
D016232:Endothelins; D011956:Receptors, Cell Surface; D017466:Receptors, Endothelin; D004247:DNA; D019204:GTP-Binding Proteins; D002118:Calcium