How the Mongoose Can Fight the Snake: The Binding Site of the Mongoose Acetylcholine Receptor

Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7717-21. doi: 10.1073/pnas.89.16.7717.

Abstract

The ligand binding site of the nicotinic acetylcholine receptor (AcChoR) is within a short peptide from the alpha subunit that includes the tandem cysteine residues at positions 192 and 193. To elucidate the molecular basis of the binding properties of the AcChoR, we chose to study nonclassical muscle AcChoRs from animals that are resistant to alpha-neurotoxins. We have previously reported that the resistance of snake AcChoR to alpha-bungarotoxin (alpha-BTX) may be accounted for by several major substitutions in the ligand binding site of the receptor. In the present study, we have analyzed the binding site of AcChoR from the mongoose, which is also resistant to alpha-neurotoxins. It was shown that mongoose AcChoR does not bind alpha-BTX in vivo or in vitro. cDNA fragments of the alpha subunit of mongoose AcChoR corresponding to codons 122-205 and including the presumed ligand binding site were cloned, sequenced, and expressed in Escherichia coli. The expressed protein fragments of the mongoose, as well as of snake receptors, do not bind alpha-BTX. The mongoose fragment is highly homologous (greater than 90%) to the respective mouse fragment. Out of the seven amino acid differences between the mongoose and mouse in this region, five cluster in the presumed ligand binding site, close to cysteines 192 and 193. These changes are at positions 187 (Trp----Asn), 189 (Phe----Thr), 191 (Ser----Ala), 194 (Pro----Leu), and 197 (Pro----His). The mongoose like the snake AcChoR has a potential glycosylation site in the binding site domain. Sequence comparison between species suggests that substitutions at positions 187, 189, and 194 are important in determining the resistance of mongoose and snake AcChoR to alpha-BTX. In addition, it was shown that amino acid residues that had been reported to be necessary for acetylcholine binding are conserved in the toxin-resistant animals as well.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Binding Sites
  • Blotting, Northern
  • Bungarotoxins / metabolism
  • Cloning, Molecular
  • DNA / genetics
  • DNA / isolation & purification
  • Herpestidae / genetics*
  • Humans
  • Mice
  • Molecular Sequence Data
  • Oligodeoxyribonucleotides
  • Poly A / genetics
  • Poly A / isolation & purification
  • RNA / genetics
  • RNA / isolation & purification
  • RNA, Messenger
  • Rabbits
  • Receptors, Nicotinic / genetics*
  • Receptors, Nicotinic / metabolism
  • Sequence Homology, Nucleic Acid
  • Snakes / genetics
  • Species Specificity

Substances

  • Bungarotoxins
  • Oligodeoxyribonucleotides
  • RNA, Messenger
  • Receptors, Nicotinic
  • Poly A
  • RNA
  • DNA

Associated data

  • GENBANK/M93639