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Molecular cloning and expression of the cDNA for the alpha 1A-adrenergic receptor. The gene for which is located on human chromosome 5

Pharmacological and molecular cloning studies have demonstrated heterogeneity of alpha 1-adrenergic receptors. We have now cloned two alpha 1-adrenergic receptors from a rat cerebral cortex cDNA library, using the hamster alpha 1B-adrenergic receptor as a probe. The deduced amino acid sequence of cl...

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Published in:The Journal of biological chemistry 1991-04, Vol.266 (10), p.6365-6369
Main Authors: Lomasney, J W, Cotecchia, S, Lorenz, W, Leung, W Y, Schwinn, D A, Yang-Feng, T L, Brownstein, M, Lefkowitz, R J, Caron, M G
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cited_by cdi_FETCH-LOGICAL-c433t-19bdb3341d62c12d6b21e5d12df977c14d592b22bf37826bdea357668072240b3
cites cdi_FETCH-LOGICAL-c433t-19bdb3341d62c12d6b21e5d12df977c14d592b22bf37826bdea357668072240b3
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container_title The Journal of biological chemistry
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creator Lomasney, J W
Cotecchia, S
Lorenz, W
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Brownstein, M
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Caron, M G
description Pharmacological and molecular cloning studies have demonstrated heterogeneity of alpha 1-adrenergic receptors. We have now cloned two alpha 1-adrenergic receptors from a rat cerebral cortex cDNA library, using the hamster alpha 1B-adrenergic receptor as a probe. The deduced amino acid sequence of clone RA42 encodes a protein of 560 amino acids whose putative topology is similar to that of the family of G-protein-coupled receptors. The primary structure though most closely resembles that of an alpha 1-adrenergic receptor, having approximately 73% amino acid identity in the putative transmembrane domains with the previously isolated hamster alpha 1B receptor. Analysis of the ligand binding properties of RA42 expressed in COS-7 cells with a variety of adrenergic ligands demonstrates a unique alpha 1-adrenergic receptor pharmacology. High affinity for the antagonist WB4101 and agonists phenylephrine and methoxamine suggests that cDNA RA42 encodes the alpha 1A receptor subtype. Northern blot analysis of various rat tissues also shows the distribution expected of the alpha 1A receptor subtype with abundant expression in vas deferens followed by hippocampus, cerebral cortex, aorta, brainstem, heart and spleen. The second alpha 1-adrenergic receptor cloned represents the rat homolog of the hamster alpha 1B subtype. Expression of mRNA for this receptor is strongly detected in liver followed by heart, cerebral cortex, brain stem, kidney, lung, and spleen. This study provides definitive evidence for the existence of three alpha 1-adrenergic receptor subtypes.
doi_str_mv 10.1016/S0021-9258(18)38126-2
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The primary structure though most closely resembles that of an alpha 1-adrenergic receptor, having approximately 73% amino acid identity in the putative transmembrane domains with the previously isolated hamster alpha 1B receptor. Analysis of the ligand binding properties of RA42 expressed in COS-7 cells with a variety of adrenergic ligands demonstrates a unique alpha 1-adrenergic receptor pharmacology. High affinity for the antagonist WB4101 and agonists phenylephrine and methoxamine suggests that cDNA RA42 encodes the alpha 1A receptor subtype. Northern blot analysis of various rat tissues also shows the distribution expected of the alpha 1A receptor subtype with abundant expression in vas deferens followed by hippocampus, cerebral cortex, aorta, brainstem, heart and spleen. The second alpha 1-adrenergic receptor cloned represents the rat homolog of the hamster alpha 1B subtype. 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ispartof The Journal of biological chemistry, 1991-04, Vol.266 (10), p.6365-6369
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subjects Amino Acid Sequence
Animals
Autoradiography
Base Sequence
Blotting, Northern
Blotting, Southern
Chromosome Mapping
Chromosomes, Human, Pair 5
Cloning, Molecular
DNA - genetics
Gene Expression Regulation
Humans
Molecular Sequence Data
Plasmids
Rats
Receptors, Adrenergic, alpha - genetics
Restriction Mapping
RNA - analysis
title Molecular cloning and expression of the cDNA for the alpha 1A-adrenergic receptor. The gene for which is located on human chromosome 5
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