Solubilization of sodium channel from human brain.
[3H]Tetrodotoxin binds to a single class of receptor sites in homogenates of human brain with a KD of 9.1 nM at 0 degree 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.
de Rycker
C
C
Laboratoire de Biochimie Générale et Comparée, Université de Liège, Belgium.
Grandfils
C
C
Bettendorff
L
L
Schoffeniels
E
E
D000328:Adult; D000818:Animals; D001921:Brain / Q000378:metabolism* / Q000648:ultrastructure; D002122:Calcium Chloride / Q000494:pharmacology; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002793:Cholic Acids; D003902:Detergents; D004557:Electric Organ / Q000378:metabolism; D004593:Electrophorus / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008297:Male; D017830:Octoxynol; D010743:Phospholipids / Q000494:pharmacology; D011092:Polyethylene Glycols; D015222:Sodium Channels / Q000378:metabolism*; D012995:Solubility; D013779:Tetrodotoxin / Q000378:metabolism
D002352:Carrier Proteins; D002793:Cholic Acids; D003902:Detergents; D010743:Phospholipids; D015222:Sodium Channels; C023790:tetrodotoxin-binding protein; D011092:Polyethylene Glycols; D013779:Tetrodotoxin; D017830:Octoxynol; C010615:Nonidet P-40; D002122:Calcium Chloride; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate