Cloning and expression of a human endothelin receptor: subtype A
| Autor: | Jean S. Lynch, Patricia M. Rose, Bernadette K. Kienzle, Maria L. Webb, Marschall S. Runge, Edward C.-K. Liu, David J. Hayzer, Eleanor Brinson, Elizabeth A. Bogosian |
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| Rok vydání: | 1992 |
| Předmět: |
Signal peptide
Placenta Molecular Sequence Data Biology Molecular cloning Transfection Polymerase Chain Reaction Muscle Smooth Vascular Cell Line Pregnancy Complementary DNA Sequence Homology Nucleic Acid Animals Humans Amino Acid Sequence Cloning Molecular Receptor Aorta Cells Cultured Gene Library chemistry.chemical_classification Base Sequence Sequence Homology Amino Acid cDNA library Receptors Endothelin Endothelins Cell Membrane General Medicine Molecular biology Amino acid Rats Open reading frame Transmembrane domain Kinetics Biochemistry chemistry Oligodeoxyribonucleotides Cattle Female |
| Zdroj: | The American journal of the medical sciences. 304(4) |
| ISSN: | 0002-9629 |
| Popis: | The polymerase chain reaction, employing degenerate primers specific for the intramembrane domains III and VI of G-coupled receptors, was used to generate partial clones encoding those receptors carried by cultured rat aorta smooth muscle cells. One clone, spanning the intramembrane domains IV–VI of a receptor specific for endothelin-1 (ET-R[A]), was used as a probe to screen a human placental cDNA library. The clone pL4-3, encoding a selective type of human endothelin receptor (ET-R[A]), has an open reading frame encoding a protein 427 amino acids in length, with a relative molecular weight of 48, 625 daltons. The sequence analysis suggests the presence of a signal peptide, two potential sites for glycosylation in the N terminal extracellular domain, the seven transmembrane domains typical of G-protein receptors, and several potential sites for phosphorylation in the C terminal cytoplasmic domain. At the amino acid level, the human ET-R(A) shows 91% and 94% identity with the rat and bovine ET-R(A)s, respectively, and 59% similarity with the human ET-R(B). Xenopus laevis oocytes injected with the cloned cDNA express binding sites specific for endothelin-1. Expression of the message in COS 7 cells gave a membrane-bound product to which binding of the [125I]-ET-1 was inhibited by peptide analogues specific for ET-R(A). |
| Databáze: | OpenAIRE |
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