[Purification of sodium channel from squid mantle and reincorporation in planar lipid bilayers].
The purification of voltage-dependent sodium channels from mantle muscles of the squid is monitored through increase in the specific binding of tritiated saxitoxin (3H-STX) of membrane extracts solubilized with Lubrol-PX, chromatographed over an ion-exchange resin then through an affinity column coupling lectins (WGA) and Sepharose. The achieved purification factor is 300 and the electrophoretic pattern of purified fractions shows a single band at 260 kD. Reincorporation of these fractions into planar lipid bilayers yields two conductance levels (in 0.5 M NaCl). The most frequently observed level, at 15 pS, is in agreement with in situ recordings. The other level, at 150 pS, equally blocked by tetrodotoxin, could be explained by some flexibility of the open state or alternatively by cooperative interactions between channels.
Bendahhou
S
S
Laboratoire Polymères, Biopolymères, Membranes, U.R.A.-C.N.R.S., n.500, Faculté des Sciences et Techniques, Mont-Saint-Aignan.
Thoiron
B
B
Duclohier
H
H
D000818:Animals; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D049832:Decapodiformes / Q000033:anatomy & histology*; D004591:Electrophoresis, Polyacrylamide Gel; D008051:Lipid Bilayers; D009132:Muscles / Q000737:chemistry*; D015222:Sodium Channels / Q000737:chemistry*
D008051:Lipid Bilayers; D015222:Sodium Channels
Purification du canal sodique du manteau de calmar et réincorporation dans des bicouches lipidiques planes.