Isolation of two saxitoxin-sensitive sodium channel subtypes from rat brain with distinct biochemical and functional properties.
Two different 3H-saxitoxin-binding proteins, with distinct biochemical and functional properties, were isolated from rat brain using a combination of anion exchange and lectin affinity chromatography as well as high resolution size exclusion and anion exchange HPLC. The alpha subunits of the binding proteins had different apparent molecular weights on SDS-PAGE (Type A: 235,000; Type B: 260,000). When reconstituted into planar lipid bilayers, the two saxitoxin-binding proteins formed sodium channels with different apparent single-channel conductances in the presence of batrachotoxin (Type A: 22 pS; Type B: 12 pS) and veratridine (Type A: 9 pS; Type B: 5 pS). The subtypes were further distinguished by scorpion (Leiurus quinquestriatus) venom which had different effects on single-channel conductance and gating of veratridine-activated Type A and Type B channels. Scorpion venom caused a 19% increase in single-channel conductance of Type A channels and a 35-mV hyperpolarizing shift in activation. Scorpion venom doubled the single-channel conductance of Type B channels and shifted activation by at least 85 mV.
Corbett
A M
AM
Department of Physiology, University of Maryland School of Medicine, Baltimore 21201.
Krueger
B K
BK
D029845:Amphibian Proteins; D000818:Animals; D001496:Batrachotoxins / Q000494:pharmacology; D001921:Brain / Q000378:metabolism*; D002352:Carrier Proteins / Q000302:isolation & purification* / Q000378:metabolism; D002845:Chromatography; D004553:Electric Conductivity; D051381:Rats; D012530:Saxitoxin / Q000378:metabolism* / Q000494:pharmacology; D012604:Scorpion Venoms / Q000494:pharmacology; D015222:Sodium Channels / Q000378:metabolism*; D014701:Veratridine / Q000494:pharmacology
D029845:Amphibian Proteins; D001496:Batrachotoxins; D002352:Carrier Proteins; D012604:Scorpion Venoms; D015222:Sodium Channels; C027525:saxitoxin-binding protein, Rana catesbeiana; D012530:Saxitoxin; D014701:Veratridine