Photoaffinity labeling of the tetrodotoxin binding component of Electrophorus electricus electroplax.

Abstract:

:Radioactive azide derivatives of tetrodotoxin (TTX) were synthesized using 2-nitro-4-azidephenyl-[3H]beta 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. N3 showed reversible binding to membranes of Electrophorus electricus electroplax in the dark with Kd = 30 nM and B max = 5.2 pmol/mg protein. By photoirradiation, irreversible binding of N3 to the membranes was observed. 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.

journal_name

Neurochem Res

journal_title

Neurochemical research

authors

Uehara S,Uyemura K

doi

10.1007/BF00965886

subject

Has Abstract

pub_date

1985-08-01 00:00:00

pages

1119-28

issue

8

eissn

0364-3190

issn

1573-6903

journal_volume

10

pub_type

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