Molecular cloning and pharmacological characterization of bovine calcitonin repeptor-like receptor from bovine aortic endothelial cells

被引：14

作者：

Aiyar, N ^{[1
]}

Disa, J ^{[1
]}

Ao, ZH ^{[1
]}

Xu, D ^{[1
]}

Surya, A ^{[1
]}

Pillarisetti, K ^{[1
]}

Parameswaran, N ^{[1
]}

Gupta, SK ^{[1
]}

Douglas, SA ^{[1
]}

Nambi, P ^{[1
]}

机构：

[1] GlaxoSmithKline, Dept Cardiovasc Pharmacol, King Of Prussia, PA 19406 USA

来源：

BIOCHEMICAL PHARMACOLOGY | 2002年 / 63卷 / 11期

关键词：

adrenomedulin; calcitonin gene-related peptide; bovine calcitonin receptor-like receptor; receptor activity modifying protein; radioligand binding; adenylyl cyclase;

D O I：

10.1016/S0006-2952(02)00990-5

中图分类号：

R9 [药学];

学科分类号：

1007 ;

摘要：

A complementary DNA encoding calcitonin receptor-like receptor (CRLR) was isolated from a bovine aortic endothelial cell library. The bovine CRLR has 462 amino acids and 92% homology with the human CRLR. In a reverse transcriptase-polymerase chain reaction assay, bovine CRLR was found to be widely distributed, including in the heart and lungs. Stable transfection of bovine CRLR in human embryonic kidney cells (HEK-293) resulted in specific high-affinity [I-125] rat adrenomedulin (rADM)-binding (dissociation constant = 145 +/- 15 pM). ADM-stimulated adenylyl cyclase activity with an EC50 value, of 5.0 +/- 1.2 nM. The human ADM receptor antagonist hADM(22-52) inhibited [I-125]rADM-binding and ADM-stimulated adenylyl cyclase activity. Interactions between bovine CRLR and individual receptor activity modifying proteins (RAMPs) were also investigated. Transient co-transfection of bovine CRLR cDNA with human receptor activity modifying protein 1 (hRAMP1) cDNA in HEK-293 cells resulted in the expression of a CRLR that displayed high-affinity binding to calcitonin gene-related peptide. Co--transfection of bovine CRLR with human RAMP2 or RAMP3 cDNAs in HEK-293 cells displayed high-affinity ADM receptors. These observations suggest that in the absence of exogenous RAMPs heterologous expression of bovine CRLR results in an ADM receptor phenotype. (C) 2002 Elsevier Science Inc. All rights reserved.

引用

页码：1949 / 1959

页数：11

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