Mapping the functional topography of a receptor

Bioorg Khim. 1992 Oct-Nov;18(10-11):1319-29.

Abstract

Photoactivatable derivatives of the alpha-neurotoxin II from Naja naja oxiana are useful tools for investigating the three dimensional architecture of the extra-membrane part of the nicotinic acetylcholine receptor from the electric tissue of Torpedo californica. Three derivatives, carrying an azidobenzoyl group in position Lys-15, Lys-26, and Lys-46, respectively, are shown to react differently within the receptor's quaternary structure. Especially the Lys-26 and Lys-46 derivatives can be used for differentiating between the two nonequivalent alpha-subunits. The Lys-26 derivative is applied for probing the receptor subunits next to the alpha-subunit: the gamma-subunit is shown to be located next to the alpha-subunit binding d-tubocurarine with high affinity. The delta-subunit is the neighbor of the low affinity alpha-subunit. We radioiodinated the toxin derivatives and localized the 125I at the His-31 residue of the toxin. Very little label was found in position Tyr-24, the only tyrosine residue of the toxin, or in position His-4, the only other histidine residue. This result is important for the cleavage experiments necessary in attempts to identify the receptor sequence which reacted with the photolabel.

MeSH terms

  • Affinity Labels
  • Amino Acid Sequence
  • Animals
  • Chromatography, Liquid
  • Cobra Neurotoxin Proteins / chemistry
  • Cross-Linking Reagents
  • Electrophoresis, Polyacrylamide Gel
  • Molecular Sequence Data
  • Photochemistry
  • Receptors, Nicotinic / chemistry*
  • Receptors, Nicotinic / physiology
  • Snakes
  • Torpedo

Substances

  • Affinity Labels
  • Cobra Neurotoxin Proteins
  • Cross-Linking Reagents
  • Receptors, Nicotinic
  • neurotoxin II, Naja naja oxiana