Preparation of [125I-Tyr27,Leu5]beta h-endorphin and its use for crosslinking of opioid binding sites in human striatum and NG108-15 neuroblastoma-glioma cells.
A radioligand suitable for crosslinking studies to opioid receptors has been obtained by radioiodination and purification of the monoiodotyrosine-27 derivative of the synthetic human beta-endorphin (beta h-endorphin) analogue [5-leucine]beta h-endorphin. The derivative, [27-[125I]monoiodotyrosine,5-leucine]beta h-endorphin, was crosslinked to human striatal (caudate and putamen) and NG108-15 neuroblastoma-glioma cell membranes by using disuccinimidyl suberate. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis under reducing conditions revealed four specifically labeled bands at 68, 40, 30, and 25 kDa for both human caudate and putamen, whereas NG108-15 cell membranes gave specifically labeled bands at 92, 56, 38, and 23 kDa.
D000328:Adult; D000818:Animals; D002421:Caudate Nucleus / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003432:Cross-Linking Reagents; D004723:Endorphins / Q000138:chemical synthesis* / Q000378:metabolism; D005910:Glioma / Q000378:metabolism; D006801:Humans; D008297:Male; D051379:Mice; D008970:Molecular Weight; D009447:Neuroblastoma / Q000378:metabolism; D011699:Putamen / Q000378:metabolism*; D051381:Rats; D011957:Receptors, Opioid / Q000378:metabolism*
D003432:Cross-Linking Reagents; D004723:Endorphins; D011957:Receptors, Opioid