How do short neurotoxins bind to a muscular-type nicotinic acetylcholine receptor?
We investigated the interacting surface between a short curarimimetic toxin and a muscular-type nicotinic acetylcholine receptor, looking for the ability of various biotinylated Naja nigricollis alpha-neurotoxin analogues to bind simultaneously the receptor and streptavidin. All these derivatives, modified at positions 10 (loop I), 27, 30, 33, 35 (loop II), 46, and 47 (loop III) or the N-terminal (erabutoxin numbering), still shared high affinity for the receptor, and in the absence of receptor they all bound soluble streptavidin. However, the proportion of the toxin-receptor complex that bound to streptavidin-coated beads, varied both with the location of the modification and with the length of the linker between biotin and the toxin. In the receptor-toxin complex, the concave side of loops II and III was not accessible to streptavidin, unlike the N terminus of the toxin and, to a certain extent, loop I. On the convex face, loop III was the most accessible, whereas the tip of loop II, especially Arg-30, seemed to be closer to the receptor. The present data demonstrate that short toxins neither penetrate deeply into a crevice as proposed earlier nor lie parallel to the receptor extracellular wall. These data also suggest that they may not lie strictly perpendicular to the cylindrical wall of the receptor. These results fit nicely with three-dimensional models of interaction between long neurotoxins and their receptors and support the idea that short and long curarimimetic toxins share a similar overall topology of interaction when bound to nicotinic receptors.
Teixeira-Clerc
Fatima
F
Commissariat à l'Energie Atomique, CEA/Saclay, Département d'Ingénierie et d'Etudes des Protéines, Gif-sur-Yvette 91191, France.
Ménez
André
A
Kessler
Pascal
P
D000818:Animals; D001710:Biotin / Q000378:metabolism; D008958:Models, Molecular; D009498:Neurotoxins / Q000378:metabolism*; D011485:Protein Binding; D011978:Receptors, Nicotinic / Q000378:metabolism*; D019809:Streptavidin / Q000378:metabolism; D014101:Torpedo
D009498:Neurotoxins; D011978:Receptors, Nicotinic; D001710:Biotin; D019809:Streptavidin