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January 18, 1983Biochemistry

Principal glycopeptide of the tetrodotoxin/saxitoxin binding protein from Electrophorus electricus: isolation and partial chemical and physical characterization

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Authors

JMJames A. MillerQuidel Corporation (United States)WAWilliam S. AgnewValve (United States)SLS. Rock LevinsonStanford University

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Implication

Biochemical analysis demonstrates isolation of the electric eel sodium channel glycopeptide, indicating the neurotoxin binding site resides on a single high-molecular-weight subunit.

Key Points

  • To isolate and characterize the physical and chemical properties of the primary tetrodotoxin- and saxitoxin-binding polypeptide from the electroplax of Electrophorus electricus.
  • Isolated the voltage-sensitive sodium channel protein from Electrophorus electricus electroplax tissue.
  • Developed a rapid purification protocol to prepare milligram quantities of the pure, sodium dodecyl sulfate (SDS)-denatured polypeptide.
  • Analyzed the amino acid and carbohydrate compositions and evaluated electrophoretic behavior using SDS-polyacrylamide gel electrophoresis.
  • Obtained homogeneous preparations exhibiting high specific activity (≥ 2000 pmol of TTX binding sites/mg of protein) consisting exclusively of the large channel-forming polypeptide.
  • Demonstrated that the true molecular weight likely exceeds the earlier estimate of 260,000 Da due to anomalous electrophoretic mobility driven by high SDS binding and bulky carbohydrate substituents.

Cite This Study

Miller et al. (1983) studied this question.

synapsesocial.com/papers/6a762dff74b63c188eb222fbhttps://doi.org/10.1021/bi00271a032
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Also Consider

Synapse has enriched 5 closely related papers on similar clinical questions. Consider them for comparative context:

  1. 1Covalent labeling of protein components of the sodium channel with a photoactivable derivative of scorpion toxin.1980 · 202 citations
  2. 2Purification of the saxitoxin receptor of the sodium channel from rat brain.1981 · 223 citations
  3. 3Immunocytochemical localization of sodium channel distributions in the excitable membranes of Electrophorus electricus.1982 · 102 citations
  4. 4Solubilized tetrodotoxin binding component from the electroplax of Electrophorus electricus. Stability as a function of mixed lipid-detergent micelle composition1979 · 82 citations
  5. 5Electron microscopic visualization of the tetrodotoxin-binding protein from Electrophorus electricus.1982 · 22 citations