A photoaffinity probe for the polyamine site regulating the NMDA receptor.
Polyamines produce on the NMDA-receptor channel activity a regulatory effect subsequent to their binding to specific sites on the receptor-complex. The photoactivatable polyamine derivative L-azidophenylspermine shows properties which suggest its potential as a photoaffinity probe to investigate the nature and topology of these sites. In the dark, its effect on the binding of tritiated N-(1-[thienyl]cyclohexyl)piperidine ([3H]TCP) to synaptosomal plasma membranes is similar to that of diaminodecane. Arcaine antagonizes both the effects of L-azidophenylspermine and diaminodecane on [3H]TCP binding. L-Azidophenylspermine competes in post-synaptic densities with tritiated spermidine for a unique binding site with an EC50 similar to that of spermine. Upon irradiation, L-azidophenyl-spermine incorporates into this material with a high efficiency to a level consistent with both the Bmax for tritiated spermidine and the estimated density of NMDA receptors in this fraction.
Foucaud
B
B
Laboratoire de Chimie Bioorganique, CNRS URA 1386, Université Louis Pasteur, School of Pharmacy, Illkirch, France.
Gambs
S
S
Schmid
N
N
Behr
J P
JP
Goeldner
M
M
D000345:Affinity Labels; D000818:Animals; D001386:Azides / Q000737:chemistry; D015317:Biogenic Polyamines / Q000378:metabolism*; D066298:In Vitro Techniques; D010622:Phencyclidine / Q000031:analogs & derivatives / Q000378:metabolism; D010777:Photochemistry; D051381:Rats; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism*; D013095:Spermidine / Q000378:metabolism; D013096:Spermine / Q000031:analogs & derivatives / Q000737:chemistry; D013570:Synaptic Membranes / Q000378:metabolism; D013574:Synaptosomes / Q000378:metabolism
D000345:Affinity Labels; D001386:Azides; D015317:Biogenic Polyamines; D016194:Receptors, N-Methyl-D-Aspartate; C096004:azidophenylspermine; D013096:Spermine; C012058:tenocyclidine; D010622:Phencyclidine; D013095:Spermidine