The binding of BmK abT, a unique neurotoxin, to mammal brain and insect Na + channels using biosensor

Abstract

The binding properties of BmK abT (a novel neurotoxic polypeptide abT from Chinese scorpion Buthus martensi Karsch), a unique neurotoxin from Chinese scorpion, on mammal brain and insect sodium channels were investigated using the BIAcore assay. Results showed that BmK abT could bind to rat brain synaptosomes with an association rate constant of about 2.49×10 6 M −1 s −1 and a dissociation rate constant of about 1.57×10 −4 s −1, and to Heliothis nerve cord synaptosomes with an association rate constant of about 1.21×10 7 M −1 s −1 and a dissociation rate constant of about 0.99×10 −3 s −1. The binding of BmK abT to rat brain synaptosomes could be partially inhibited by increasing the membrane potential, but not by BmK AS (a novel active polypeptide AS from B. martensi Karsch), BmK IT2 (a depressant insect-selective toxin IT2 from B. martensi Karsch), and BmK I (an α-like anti-mammal toxin I from B. martensi Karsch). Binding was not modulated by veratridine. In addition, the binding of BmK abT to Heliothis nerve cord synaptosomes was significantly enhanced by increasing the membrane potential and veratridine concentration and was inhibited by BmK IT2, but not by BmK AS or BmK I. The results suggest that BmK abT binds to a distinct receptor site on mammal brain Na + channels and associates with a related site for depressant insect-selective toxins on insect sodium channels.