Photoaffinity Labeling of the DDT1 MF-2 Cell Alpha 1-adrenergic Receptor

Overview

Journal Mol Cell Biochem

Publisher Springer

Specialty Biochemistry

Date 1985 May 1

PMID 2862576

Citations 2

Authors

L E Cornett

J S Norris

Affiliations

Soon will be listed here.

Abstract

In this study, we have used an alpha 1-adrenergic receptor photoaffinity ligand, 2-[4-(4-azido-3-iodo-benzoyl)-piperazin-1-yl]-4-amino-6, 7-dimethoxyquinazoline (125I-APD), to label covalently the alpha 1-adrenergic receptor in a smooth muscle cell line. Our results indicate that in the absence of light, (125I)APD binds reversibly to a site in the DDT1 MF-2 cell membranes having pharmacological characteristics of an alpha 1-adrenergic receptor. Following incorporation of (125I)APD into partially purified membranes a single labeled band of protein with a Mr of 81 000 was visualized by autoradiography following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Incorporation of (125I)-APD into this band was affected by adrenergic agonists and antagonists in a manner consistent with an alpha 1-adrenergic interaction. Prazosin (alpha 1-selective) blocked incorporation of the label into the Mr = 81 000 protein while yohimbine (alpha 2-selective) did not. Of the adrenergic agonists, (-)-epinephrine and (-)-norepinephrine but not (-)-isoproterenol blocked labeling of the Mr = 81 000 protein. We conclude that the ligand binding site of the DDT1 MF-2 cell alpha 1-adrenergic receptor resides in a Mr = 81 000 protein.

Citing Articles

The oligosaccharide component of alpha 1-adrenergic receptors from BC3H1 and DDT1 muscle cells. Studies with glycosidases and photoaffinity labelling of intact cells.

Terman B, Reece J, Brown R, Insel P Biochem J. 1988; 253(2):363-70.

PMID: 2845917 PMC: 1149307. DOI: 10.1042/bj2530363.

A mechanism for testosterone modulation of alpha-1 adrenergic receptor expression in the DDT1 MF-2 smooth muscle myocyte.

Phillippe M, Saunders T, Bangalore S Mol Cell Biochem. 1991; 100(1):79-90.

PMID: 1646954 DOI: 10.1007/BF00230812.

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