Localization of azidophencyclidine-binding site on the nicotinic acetylcholine receptor alpha-subunit.
Nicotinic acetylcholine receptors in receptor-rich membranes from Torpedo californica and from T. marmorata electric tissue were photolabeled with the non-competitive inhibitor [3H]azidophencyclidine. The receptor subunits were separated on SDS-polyacrylamide gels and the alpha-subunits recovered from the gel, were subjected to Staphylococcus aureus V8 protease cleavage. The proteolytic fragments were resolved by SDS-polyacrylamide gel electrophoresis and were identified on protein blots by 125I-labeled alpha-bungarotoxin binding and by staining with concanavalin A. The site of specific azidophencyclidine labeling has been localized to the V8-18 kDa fragment which binds toxin. Labeling of the V8-18 kDa fragment was observed in the absence and in the presence of carbamylcholine. This was found for both the species of Torpedo used here.
Mosckovitz
R
R
Haring
R
R
Gershoni
J M
JM
Kloog
Y
Y
Sokolovsky
M
M
D000345:Affinity Labels; D000818:Animals; D001386:Azides / Q000378:metabolism*; D003488:Cyanogen Bromide / Q000494:pharmacology; D010450:Endopeptidases / Q000378:metabolism; D046911:Macromolecular Substances; D008970:Molecular Weight; D010446:Peptide Fragments / Q000378:metabolism; D010622:Phencyclidine / Q000031:analogs & derivatives* / Q000378:metabolism; D010777:Photochemistry; D011950:Receptors, Cholinergic / Q000378:metabolism*; D012697:Serine Endopeptidases; D014101:Torpedo
C040647:3-azidophencyclidine; D000345:Affinity Labels; D001386:Azides; D046911:Macromolecular Substances; D010446:Peptide Fragments; D011950:Receptors, Cholinergic; D010450:Endopeptidases; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D010622:Phencyclidine; D003488:Cyanogen Bromide