Schweitz H, Bidard J N, Maes P, Lazdunski M
Centre de Biochimie, Centre National de la Recherche Scientifique, Université de Nice, France.
Biochemistry. 1989 Dec 12;28(25):9708-14. doi: 10.1021/bi00451a025.
A polypeptide was identified in the venom of the scorpion Leiurus quinquestriatus hebraeus by its potency to inhibit the high-affinity binding of the radiolabeled snake venom toxin dendrotoxin I (125I-DTX1) to its receptor site. It has been purified, and its properties investigated by different techniques were found to be similar to those of MCD and DTXI, two polypeptide toxins active on a voltage-dependent K+ channel. However, its amino acid sequence was determined, and it was shown that this toxin is in fact charybdotoxin (ChTX), a toxin classically used as a specific tool to block one class of Ca2+-activated K+ channels. ChTX, DTXI, and MCD are potent convulsants and are highly toxic when injected intracerebroventricularly in mice. Their toxicities correlate well with their affinities for their receptors in rat brain. These three structurally different toxins release [3H]GABA from preloaded synaptosomes, the efficiency order being DTXI greater than ChTX greater than MCD. Both binding and cross-linking experiments of ChTX to rat brain membranes and to the purified MCD/DTXI binding protein have shown that the alpha-subunit (Mr = 76K-78K) of the MCD/DTXI-sensitive K+ channel protein also contains the ChTX binding sites. Binding sites for DTXI, MCD, and ChTX are in negative allosteric interaction. Our results show that charybdotoxin belongs to the family of toxins which already includes the dendrotoxins and MCD, which are blockers of voltage-sensitive K+ channels. ChTX is clearly not selective for Ca2+-activated K+ channel.
通过抑制放射性标记的蛇毒毒素树突毒素I(125I-DTX1)与其受体位点的高亲和力结合能力,在以色列金蝎毒液中鉴定出一种多肽。该多肽已被纯化,通过不同技术对其性质进行研究后发现,它与MCD和DTXI相似,这两种多肽毒素对电压依赖性钾通道有活性。然而,测定了其氨基酸序列,结果表明这种毒素实际上是蝎毒氯毒素(ChTX),一种传统上用作阻断一类钙激活钾通道的特异性工具的毒素。ChTX、DTXI和MCD都是强效惊厥剂,经脑室内注射到小鼠体内时具有高度毒性。它们的毒性与它们对大鼠脑中受体的亲和力密切相关。这三种结构不同的毒素从预先装载的突触体中释放[³H]GABA,效率顺序为DTXI>ChTX>MCD。ChTX与大鼠脑膜以及纯化的MCD/DTXI结合蛋白的结合和交联实验均表明,MCD/DTXI敏感钾通道蛋白的α亚基(Mr = 76K - 78K)也含有ChTX结合位点。DTXI、MCD和ChTX的结合位点存在负变构相互作用。我们的结果表明,蝎毒氯毒素属于已经包括树突毒素和MCD的毒素家族,它们都是电压敏感钾通道的阻断剂。ChTX显然对钙激活钾通道没有选择性。