Brain alpha-bungarotoxin binding protein cDNAs and MAbs reveal subtypes of this branch of the ligand-gated ion channel gene superfamily.
alpha-Bungarotoxin (alpha Bgt) is a potent, high-affinity antagonist for nicotinic acetylcholine receptors (AChRs) from muscle, but not for AChRs from neurons. Both muscle and neuronal AChRs are thought to be formed from multiple homologous subunits aligned around a central cation channel whose opening is regulated by ACh binding. In contrast, the exact structure and function of high-affinity alpha Bgt binding proteins (alpha BgtBPs) found in avian and mammalian neurons remain unknown. Here we show that cDNA clones encoding alpha BgtBP alpha 1 and alpha 2 subunits define alpha BgtBPs as members of a gene family within the ligand-gated ion channel gene superfamily, but distinct from the gene families of AChRs from muscles and nerves. Subunit-specific monoclonal antibodies raised against bacterially expressed alpha BgtBP alpha 1 and alpha 2 subunit fragments reveal the existence of at least two different alpha BgtBP subtypes in embryonic day 18 chicken brains. More than 75% of all alpha BgtBPs have the alpha 1 subunit, but no alpha 2 subunit, and a minor alpha BgtBP subtype (approximately 15%) has both the alpha 1 and alpha 2 subunits.
Schoepfer
R
R
Receptor Biology Laboratory, Salk Institute for Biological Studies, San Diego, California 92138-9216.
Conroy
W G
WG
Whiting
P
P
Gore
M
M
Lindstrom
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D001419:Bacteria / Q000378:metabolism; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D002038:Bungarotoxins / Q000378:metabolism; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D005796:Genes; D015640:Ion Channel Gating / Q000235:genetics*; D008024:Ligands; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D011950:Receptors, Cholinergic / Q000235:genetics* / Q000276:immunology; D011978:Receptors, Nicotinic; D011994:Recombinant Proteins; D064569:alpha7 Nicotinic Acetylcholine Receptor
D000911:Antibodies, Monoclonal; D002038:Bungarotoxins; D008024:Ligands; D010446:Peptide Fragments; D011950:Receptors, Cholinergic; D011978:Receptors, Nicotinic; D011994:Recombinant Proteins; D064569:alpha7 Nicotinic Acetylcholine Receptor; D004247:DNA