Neumann_1985_Proc.Natl.Acad.Sci.U.S.A_82_3490

Reference

Title : Antibodies to synthetic peptides as probes for the binding site on the alpha subunit of the acetylcholine receptor - Neumann_1985_Proc.Natl.Acad.Sci.U.S.A_82_3490
Author(s) : Neumann D , Gershoni JM , Fridkin M , Fuchs S
Ref : Proc Natl Acad Sci U S A , 82 :3490 , 1985
Abstract :

Synthetic peptides and their respective antibodies were used in an attempt to localize and identify the ligand-binding site of the nicotinic acetylcholine receptor. Two peptides of the receptor alpha subunit were synthesized, the first corresponding to the NH2-terminal domain (positions 1-20) and the other, to a segment (residues 126-143) that contains the first two cysteine residues. Specific antipeptide antibodies were elicited in rabbits after immunization with the peptides conjugated to bovine serum albumin. The antipeptide antibodies thus obtained cross-reacted with the receptor and bound specifically to its alpha subunit. The antipeptide antibodies were used to test whether the peptide sequences corresponded to the alpha-bungarotoxin (alpha-BTX)-binding site. Staphylococcus aureus V8-protease digestion of the isolated receptor alpha subunit generated several fragments. Antipeptide (1-20) and antipeptide (126-143) both bound a 26-kDa fragment, whereas only antipeptide (126-143) bound a 17-kDa fragment. None of these fragments were found to bind alpha-BTX. On the other hand, alpha-BTX bound to an 18-kDa fragment that did not react with either of the antipeptide antibodies. Moreover, the 26-kDa and 17-kDa fragments were also found to contain the endoglycosidase H-susceptible oligosaccharide chain. Our results indicate that the toxin-binding site lies beyond the first possible V8 protease cleavage site after residues 126-143: i.e., Asp-152. This location is in agreement with the possibility that cysteine residues 192 and/or 193 are in close proximity to or contiguous with the ligand-binding site.

PubMedSearch : Neumann_1985_Proc.Natl.Acad.Sci.U.S.A_82_3490
PubMedID: 2582416

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Citations formats

Neumann D, Gershoni JM, Fridkin M, Fuchs S (1985)
Antibodies to synthetic peptides as probes for the binding site on the alpha subunit of the acetylcholine receptor
Proc Natl Acad Sci U S A 82 :3490

Neumann D, Gershoni JM, Fridkin M, Fuchs S (1985)
Proc Natl Acad Sci U S A 82 :3490