(2-sulfonatoethyl)methanethiosulfonate and Neurodegenerative-Diseases

BNC1 is a mammalian neuronal cation channel in the novel DEG/ENaC ion channel family. BNC1 channels are transiently activated by extracellular protons and are constitutively activated by insertion of large residues, such as valine, in place of Gly-430; residue 430 is a site where analogous mutations in some Caenorhabditis elegans family members cause a swelling neurodegeneration. Mutation of Gly-430 to a small amino acid, cysteine, neither generated constitutive currents nor allowed modification of this residue by sulfhydryl-reactive methanethiosulfonate (MTS) compounds. However, when protons activated the channel, Cys-430 became accessible to extracellular MTS reagents, which modified Cys-430 to generate constitutive currents. Fluorescent MTS reagents also labeled Cys-430 in activated channels. These data indicate that protons induce a reversible conformational change that activates BNC1 thereby exposing residue 430 to the extracellular solution. Once Cys-430 is modified with a large chemical group, the channel is prevented from relaxing back to the inactive state. These results link ligand-dependent activation and activation by mutations that cause neurodegeneration.

Topics: Amino Acid Substitution; Animals; Cold Temperature; Degenerin Sodium Channels; Epithelial Sodium Channels; Ethyl Methanesulfonate; Female; Indicators and Reagents; Ion Channel Gating; Ion Channels; Mesylates; Mutation; Nerve Tissue Proteins; Neurodegenerative Diseases; Neurons; Oocytes; Protein Conformation; Sodium Channels; Structure-Activity Relationship; Xenopus laevis