Photoaffinity labeling of the DDT1 MF-2 cell alpha 1-adrenergic receptor.
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.
Cornett
L E
LE
Norris
J S
JS
D000316:Adrenergic alpha-Agonists / Q000494:pharmacology; D000317:Adrenergic alpha-Antagonists / Q000494:pharmacology; D000345:Affinity Labels; D000818:Animals; D001386:Azides; D002460:Cell Line; D009130:Muscle, Smooth / Q000032:analysis*; D010777:Photochemistry; D011224:Prazosin / Q000031:analogs & derivatives / Q000494:pharmacology; D011799:Quinazolines; D011942:Receptors, Adrenergic, alpha / Q000032:analysis* / Q000187:drug effects; D015016:Yohimbine / Q000494:pharmacology
D000316:Adrenergic alpha-Agonists; D000317:Adrenergic alpha-Antagonists; D000345:Affinity Labels; D001386:Azides; D011799:Quinazolines; D011942:Receptors, Adrenergic, alpha; D015016:Yohimbine; C041552:azidoprazosin; D011224:Prazosin