Molecular Properties of neurotoxin receptors sites associated with sodium channels from mammalian brain.
Neurotoxins that act at specific receptor sites on voltage-sensitive sodium channels have been used as molecular probes to identify and purify protein components of sodium channels from mammalian brain. Photoreactive derivatives of scorpion toxin have been prepared and used to covalently label sodium channels in intact synaptosomes. Two polypeptides, alpha with Mr approximately 270,000 and beta with Mr approximately 38,000, are specifically labeled indicating that they are components of the scorpion toxin receptor site on the sodium channel. The sodium channel can be solubilized with retention of specific binding of [3H] saxitoxin using nonionic detergents such as Triton X-100. The solubilized saxitoxin receptor has molecular weight of 316,000 +/- 63,000 and binds 0.9 g of Triton X-100 and phospholipid per g of protein. The solubilized receptor can be purified 750-fold by ion exchange chromatography, wheat germ lectin/Sepharose chromatography and sucrose gradient sedimentation to a final specific activity of 1488 pmol/mg. Analysis of the polypeptide chain composition of the most highly purified fractions indicates that alpha and beta comprise 65% of the protein of these fractions and are only the polypeptides whose presence correlates with saxitoxin binding activity. These studies lead to a working hypothesis of sodium channel structure in which the intact channel is comprised of a complex with Mr of approximately 316,000 containing one mole of alpha (Mr approximately 270,000) and one to three moles of beta (Mr approximately 38,000).
D000818:Animals; D001921:Brain / Q000378:metabolism*; D007473:Ion Channels / Q000378:metabolism*; D008970:Molecular Weight; D009498:Neurotoxins / Q000378:metabolism* / Q000494:pharmacology; D011950:Receptors, Cholinergic / Q000302:isolation & purification / Q000378:metabolism*; D012910:Snake Venoms / Q000378:metabolism / Q000494:pharmacology; D012964:Sodium / Q000378:metabolism*
D007473:Ion Channels; D009498:Neurotoxins; D011950:Receptors, Cholinergic; D012910:Snake Venoms; D012964:Sodium