Differential surface accessibility of alpha(187-199) in the Torpedo acetylcholine receptor alpha subunits.
We have probed the surface accessibility of residues alpha187 to alpha199 of the Torpedo acetylcholine receptor with monoclonal antibody 383C, which binds uniquely to these residues. However, 383C binds to only one of the two alpha subunits in the membrane-bound receptor, neither of the two subunits in carbamylcholine-desensitized receptor, and to both alpha subunits in Triton X-100 solubilized receptor. The kinetics of association and dissoci-ation of 383C with the peptide alpha(183-199) compared to those with the membrane-bound receptor suggest that all but a single hydrogen bond of affinity derives from contacts between this peptide and the monoclonal antibody paratope. Inhibition of 383C binding by alpha-bungarotoxin selectively directed to the alpha subunit correlated with the high-affinity d-tubocurarine binding site, along with a lack of inhibition by alpha-bungarotoxin directed to the alpha subunit correlated with the low-affinity d-tubocurarine binding site, suggests that the 383C epitope on the membrane-bound receptor resides on the alpha subunit associated with the high-affinity d-tubocurarine binding site. The results presented here suggest a structural basis for the differences between the two receptor acetylcholine binding sites.
Fairclough
R H
RH
Department of Neurology, University of California, Davis, CA 95616, USA. rhfairclough@ucdavis.edu
Twaddle
G M
GM
Gudipati
E
E
Lin
M Y
MY
Richman
D P
DP
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001665:Binding Sites; D002038:Bungarotoxins / Q000378:metabolism; D002217:Carbachol / Q000378:metabolism; D000939:Epitopes / Q000032:analysis*; D007700:Kinetics; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D017830:Octoxynol / Q000378:metabolism; D011978:Receptors, Nicotinic / Q000737:chemistry* / Q000378:metabolism; D017186:Titrimetry; D014101:Torpedo / Q000502:physiology; D014403:Tubocurarine / Q000378:metabolism
D000911:Antibodies, Monoclonal; D002038:Bungarotoxins; D000939:Epitopes; D008565:Membrane Proteins; D011978:Receptors, Nicotinic; D002217:Carbachol; D017830:Octoxynol; D014403:Tubocurarine