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Photoaffinity labeling of the tetrodotoxin binding component ofElectrophorus electricus electroplax

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Abstract

  1. 1)

    Radioactive azide derivatives of tetrodotoxin (TTX) were synthesized using 2-nitro-4-azidephenyl-[3H]β alanine for the purpose of photolabeling of the Na channel. Three azide derivatives, N1, N2 and N3, were separated by ion exchange chromatography on Bio-Rex 70 resin and reversed phase high performance liquid chromatography. N3 was more stable and obtained at a higher yield than the other two derivatives. Bioactivity of N3 was one-twentieth of that of TTX.

  2. 2)

    N3 showed reversible binding to membranes ofElectrophorus electricus electroplax in the dark withK d=30 nM andB max=5.2 pmol/mg protein. By photoirradiation, irreversible binding of N3 to the membranes was observed.

  3. 3)

    A N3 binding component was solubilized by lubrol PX and partially purified from the electroplax membranes by Sephadex G25 and Sepharose 6B column chromatography. The component, purified 500 fold from the starting membranes, showed molecular weight of 10,000.

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Authors and Affiliations

  1. Department of Physiology, Saitama Medical School, Moroyamacho, Irumagun, 350-04, Saitama, Japan

    Seiji Uehara & Keiichi Uyemura

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  1. Seiji Uehara
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  2. Keiichi Uyemura
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Dedicated to Professor Yasuzo Tsukada

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Uehara, S., Uyemura, K. Photoaffinity labeling of the tetrodotoxin binding component ofElectrophorus electricus electroplax. Neurochem Res 10, 1119–1128 (1985). https://doi.org/10.1007/BF00965886

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  • DOI: https://doi.org/10.1007/BF00965886

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