Critical interactions at the dimer interface of kappa-bungarotoxin, a neuronal nicotinic acetylcholine receptor antagonist.
The alpha- and kappa-neurotoxins are polypeptide antagonists of nicotinic acetylcholine receptors derived from snake venom. They are structurally very similar but differ in their specificity for receptor subtype and in their native aggregation state. While the alpha-neurotoxins are monomeric, the kappa-neurotoxins occur as homodimers. The crystal structure shows that there is a correlation in the distance between essential arginine residues in the kappa-bungarotoxin dimer and the distance between the acetylcholine binding sites in the pentameric receptor. This has lead to an investigation of the critical interactions at the dimer interface of kappa-bungarotoxin. Mutations of residues that the crystal structure indicates participate in dimer interaction were found to fall into two general groups: those that do not affect the dimerization state or activity of kappa-bungarotoxin as single mutants, and those that interfere with it to such an extent that the protein is no longer able to fold properly. In general, those residues that fall into the latter group are found to be invariant in kappa-neurotoxins and not found in alpha-neurotoxins. The results suggest that the extent of both the main chain-main chain beta-sheet hydrogen bond interaction and van der Waals interactions between Phe 49 and Ile 20 are required for dimer formation. These studies provide a basis for understanding why the kappa-neurotoxins readily dimerize in solution and the alpha-neurotoxins do not and also suggest that there is a possible interrelationship between dimer formation and protein folding in kappa-bungarotoxin.
Grant
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GA
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110, USA. ggrant@pharmdec.wustl.edu
Al-Rabiee
R
R
Xu
X L
XL
Zhang
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D002038:Bungarotoxins / Q000737:chemistry* / Q000494:pharmacology*; D003001:Cloning, Molecular; D003461:Crystallography; D019281:Dimerization; D008958:Models, Molecular; D008961:Models, Structural; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009498:Neurotoxins / Q000737:chemistry*; D018733:Nicotinic Antagonists / Q000737:chemistry* / Q000494:pharmacology; D017354:Point Mutation; D017433:Protein Structure, Secondary; D011978:Receptors, Nicotinic / Q000187:drug effects*; D011994:Recombinant Proteins / Q000737:chemistry / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D012910:Snake Venoms / Q000737:chemistry*; D012911:Snakes; D012984:Software
D002038:Bungarotoxins; D009498:Neurotoxins; D018733:Nicotinic Antagonists; D011978:Receptors, Nicotinic; D011994:Recombinant Proteins; D012910:Snake Venoms