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Identification of the 5‐HT 1A Receptor Binding Subunit in Rat Brain Membranes Using the Photoaffinity Probe [ 3 H]8‐Methoxy‐2‐[ N ‐ n ‐Propyl, N ‐3‐(2‐Nitro‐4‐Azidophenyl)Aminopropyl]Aminotetralin
Author(s) -
Emerit M. B.,
Mestikawy S. El,
Gozlan H.,
Cossery J. M.,
Besselievre R.,
Marquet A.,
Hamon M.
Publication year - 1987
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1987.tb02875.x
Subject(s) - photoaffinity labeling , protein subunit , chemistry , membrane , receptor , identification (biology) , stereochemistry , biophysics , biochemistry , biology , botany , gene
The synthesis of a tritiated derivative of the 5‐HT 1A photoaffinity probe 8‐methoxy‐2‐[ N ‐ n ‐propyl, N ‐3‐(2‐nitro‐4‐azidophenyl)aminopropyl]aminotetralin ([ 3 H]8‐methoxy‐3′‐NAP‐amino‐PAT) allowed the use of this probe for attempting the irreversible labeling of specific binding sites in rat brain membranes. Sodium dodecyl‐sulfate‐polyacrylamide gel electrophoresis of proteins solubilized from hippocampal microsomal membranes that had been incubated with 20 n M [ 3 H]8‐methoxy‐3′‐NAP‐amino‐PAT under UV light revealed a marked incorporation of 3 H label into a 63‐kilodalton protein termed P I . As expected of a possible correspondence between P 1 and 5‐HT 1A receptor binding sites, 3 H labeling by the photoaffinity probe could be prevented by selective 5‐HT 1A ligands such as 8‐hydroxy‐2‐(di‐ n ‐propylamino)tetralin, ipsapirone, buspirone, and gepirone and by N‐ethylmaleimide, but not by the 5‐HT 2 antagonist ketanserin, noradrenaline‐and dopamine‐related drugs, monoamine oxidase inhibitors, and chlorimipramine. Furthermore, the regional and subcellular distributions of P 1 were identical to those of specific 5‐HT 1A binding sites. These results indicated that the binding subunit of the 5‐HT 1A receptor is a 63‐kilodalton protein with a functionally important sulfhydryl group(s).