Scanning Mutagenesis of α-Conotoxin Vc1.1 Reveals Residues Crucial for Activity at the α9α10 Nicotinic Acetylcholine Receptor

摘要: Vc1.1 is a disulfide-rich peptide inhibitor of nicotinic acetylcholine receptors that has stimulated considerable interest in these as potential therapeutic targets for the treatment neuropathic pain. Here we present an extensive series mutational studies which all residues except conserved cysteines were mutated separately to Ala, Asp, or Lys. The effect on (ACh)-evoked membrane currents at α9α10 receptor (nAChR), been implicated target alleviation pain, was then observed. analogs characterized by NMR spectroscopy determine effects mutations structure. structural fold found be preserved peptides where Pro substituted. Electrophysiological showed key functional activity are Asp5–Arg7 and Asp11–Ile15, because changes positions resulted loss nAChR. Interestingly, S4K N9A more potent than itself. A second generation mutants synthesized, namely N9G, N9I, N9L, S4R, S4K+N9A, both rat human α9/rat α10 hybrid receptor, providing mechanistic insight into involved eliciting biological function Vc1.1. most also tested α3β2, α3β4, α7 nAChR subtypes their selectivity. All selective These findings provide valuable interaction with subtype will help further development analgesic drugs.