Identification of a receptor for atrial natriuretic factor in rabbit aorta membranes by affinity cross-linking

Binding sites in rabbit aorta membranes for atrial natriuretic factor (ANF) have been specifically and covalently labeled by two methods. In the first, the photoreactive analog of ANF, 125I-azidobenzoyl-ANF, was synthesized and used to photoaffinity label ANF receptors. In the second, 125I-ANF was c...

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Veröffentlicht in:The Journal of biological chemistry 1985-09, Vol.260 (20), p.10889-10892
Hauptverfasser: Vandlen, R L, Arcuri, K E, Napier, M A
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Arcuri, K E
Napier, M A
description Binding sites in rabbit aorta membranes for atrial natriuretic factor (ANF) have been specifically and covalently labeled by two methods. In the first, the photoreactive analog of ANF, 125I-azidobenzoyl-ANF, was synthesized and used to photoaffinity label ANF receptors. In the second, 125I-ANF was covalently attached to its binding site by treatment of the 125I-ANF-receptor complex with bifunctional cross-linking agents. Analysis of the labeled proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography revealed that by both methods the same three protein bands were labeled. These bands had apparent molecular masses of 60,000, 70,000, and 120,000 daltons. With the photoaffinity label, half-maximal inhibition of labeling of each of these bands was achieved when approximately 200 pM of unlabeled ANF was included in the binding assay. These results suggest that these three different polypeptides are specific components of ANF receptors in rabbit aorta membranes.
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subjects Affinity Labels - metabolism
Animals
Aorta - metabolism
Atrial Natriuretic Factor
Azides - chemical synthesis
Azides - metabolism
Binding, Competitive
Biological and medical sciences
Cell Membrane - metabolism
Cell receptors
Cell structures and functions
Electrophoresis, Polyacrylamide Gel
Fundamental and applied biological sciences. Psychology
Kinetics
Macromolecular Substances
Molecular and cellular biology
Molecular Weight
Muscle Proteins - chemical synthesis
Muscle Proteins - metabolism
Rabbits
Receptors, Atrial Natriuretic Factor
Receptors, Cell Surface - isolation & purification
Receptors, Cell Surface - metabolism
title Identification of a receptor for atrial natriuretic factor in rabbit aorta membranes by affinity cross-linking
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