Biochemical characterization of the tetrodotoxin binding protein from Electrophorus electricus.
Biochemical characterization of the tetrodotoxin binding protein from Electrophorus electricus.
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来自电电鱼的河豚毒素结合蛋白的生化特征。
DOI：
10.1021/bi00267a029
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发表时间：
1982
期刊：
影响因子：
2.9
通讯作者：
Raftery,MA
中科院分区：
文献类型：
--
作者：
Moore,AC;Agnew,WS;Raftery,MA
安妮·C摩尔，1威廉S. Agnew，§ and Michael A.摘要：从Electrophorus electricus的洗涤剂溶解的河豚毒素结合组分的生物化学性质进行了研究，并与膜结合蛋白的那些发现进行了比较。各种非离子型去离子剂对毒素结合组分的增溶效率高，胆酸钠和脱氧胆酸钠对毒素结合组分的增溶效率低。洗涤剂溶解的制剂绑定河豚毒素和石房蛤毒素紧密和具体的，这种结合被观察到被迅速和不可逆地阻止由羧酸改性剂。由碳二亚胺和甘氨酸酯或由trimethyloxonium盐灭活可以防止河豚毒素占用的结合位点。河豚毒素在溶解制剂和膜中的结合活性被发现对各种方法具有高度抗性，这些方法被用于表征赋予神经和肌肉细胞膜电兴奋性的分子结构和机制。尽管对传播的动作电位的生物物理描述已经变得相当详细[关于综述，参见Armstrong（1975），Landowne et al.（1975），Ul-bricht（1977），and Hille（1978）]，负责的现象学通道的分子性质还没有被牢固地建立。本文报道了对一种负责动作电位早期钠电流的电压敏感性钠电导通道的生化分离和表征的连续研究结果，我们用两种高度特异性的神经毒素河豚毒素（TTX）1和石房蛤毒素（STX）作为该通道的生化定量标记。TTX和STX的特性和特异性已被广泛综述（Hille，1978;里奇和Rogart，1977）。这些分子以可逆的方式结合，相互竞争，并以高亲和力（K = 1-10 nM）结合到从细胞膜外部可接近的单一类别的位点（Narahashi等人，1967; Hille，1968，1975 a，B; Cuervo & Adelman，1970; Ulbricht &瓦格纳，1975）。显然，毒素阻断钠离子渗透途径，而不影响通道门控结构（Armstrong & Bezanilla，1973，1974; Keynes & Rojas，1974）。生理学研究数据与结合研究数据的比较提供了强有力的证据，表明毒素仅与生理学定义的钠通道结合（里奇和Rogart，1977）。来自剂量反应实验（Hille，1970）和波动分析（Sigworth，1980）的数据表明，TTX和STX与一种
Anne C. Moore, 1 William S. Agnew, § and Michael A. Raftery* abstract: Biochemical properties of a detergent-solubilized tetrodotoxin binding component from Electrophorus electricus have been examined and compared with those found for the membrane-bound protein. The toxin binding component was solubilized with high efficiency by a variety of nonionic de-tergents and with lower efficiency by sodium cholate and deoxycholate. Detergent-solubilized preparations bound tet-rodotoxin and saxitoxin tightly and specifically, and this binding was observed to be rapidly and irreversibly blocked by carboxylate-modifying reagents. Inactivation by carbodiimide and glycine ester or by a trimethyloxonium salt could be prevented by tetrodotoxin occupancy of the binding site. Tetrodotoxin binding activity in both solubilized preparations and in membranes was found to be highly resistant to pro-variety of approaches are being used to characterize the molecular structures and mechanisms which confer on the membranes of nerve and muscle cells the property of electrical excitability. Although the biophysical description of the propagated action potential has become quite detailed [for reviews, see Armstrong (1975), Landowne et al.(1975), Ul-bricht (1977), and Hille (1978)], the molecular nature of the phenomenological channels which are responsible has yet to be firmly established. We report here results from continuing studies on the biochemical isolation and characterization of a voltage-sensitive sodium conductance channel of the type responsible for the early sodium currents of the action po-tential.We have used two highly specific neurotoxins, tetrodotoxin (TTX) 1 and saxitoxin (STX) as biochemical markers for the quantitation of the channel. The properties and specificities of TTX and STX have been extensively reviewed (Hille, 1978; Ritchie & Rogart, 1977). These moleculesbind in a reversible manner, with mutual competition, and with high affinity {K¿= 1-10 nM) to a single class of sites accessible from theoutside of the cell membrane (Narahashi et al., 1967; Hille, 1968, 1975a, b; Cuervo & Adelman, 1970; Ulbricht & Wagner, 1975). Apparently the toxins block the sodium ion permeation pathway without affecting thestructures involved in channel gating (Armstrong & Bezanilla, 1973, 1974; Keynes & Rojas, 1974). Comparison of data from physiological studies with those from binding studies has provided strong evidence that the toxins bind only to the physiologically defined sodium channel (Ritchie & Rogart, 1977). Datafrom dose-response experiments (Hille, 1970) and from fluctuation analysis (Sigworth, 1980) suggest that TTX and STX bind with a
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DOI：
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发表时间：
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影响因子：
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作者：
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作者：
LEONHARDT, H;GORDON, L;LIVINGSTON, R
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