Solubilization and characterization of the insect neuronal sodium channel.
Locust neuronal sodium channels were solubilized by 1% cholate and 0.2% Triton X-100, and their functionality was monitored by [3H]saxitoxin (STX) binding assays. About 40% of STX binding activity was recovered in the solubilized fraction without affecting affinity (Kd = 0.5 nM) and the time and temperature dependent STX binding activity was significantly stabilized in the presence of 20 nM STX. Partial purification by an anion exchange resin yielded a 20% recovery and a 3.5 times increase in the specific STX binding activity. Identification of the locust solubilized sodium channels by immunoprecipitation and radiophosphorylation revealed a Mr of 245,000 on SDS-PAGE. The present solubilized preparation will enable the study of the unique pharmacology of insect sodium channels.
Moskowitz
H
H
Institute of Life Sciences, Hebrew University of Jerusalem, Israel.
Zlotkin
E
E
Gordon
D
D
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006110:Grasshoppers / Q000378:metabolism*; D007700:Kinetics; D009474:Neurons / Q000378:metabolism*; D010766:Phosphorylation; D011233:Precipitin Tests; D012530:Saxitoxin / Q000378:metabolism; D015222:Sodium Channels / Q000378:metabolism*
D015222:Sodium Channels; D012530:Saxitoxin