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Solubilization of Sodium Channel from Human Brain
Author(s) -
Rycker Cécile,
Grandfils Christian,
Bettendorff Lucien,
Schoffeniels Ernest
Publication year - 1989
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1989.tb09128.x
Subject(s) - tetrodotoxin , solubilization , phosphatidylcholine , chemistry , receptor , chaps , biophysics , sodium , membrane , chromatography , biochemistry , biology , phospholipid , organic chemistry
[ 3 H]Tetrodotoxin binds to a single class of receptor sites in homogenates of human brain with a K D of 9.1 n M at 0°C and a maximal binding capacity of 5.9 pmol/mg of protein. This tetrodotoxin receptor has been solubilized, and several parameters influencing the efficiency of this critical step have been studied. Treatment of brain membranes with 2% (wt/vol) Nonidet P‐40 solubilizes up to 38% of the tetrodotoxin receptor sites. The duration of this solubilization step must not exceed 15 min at an optimal pH of 6.8. The binding activity is most stable when exogenous phosphatidylcholine is added to the soluble receptor with a phosphatidylcholine/detergent ratio of 1:5.