Rochat H, Kharrat R, Sabatier J M, Mansuelle P, Crest M, Martin-Eauclaire M F, Sampieri F, Oughideni R, Mabrouk K, Jacquet G, Van Rietschoten J, El Ayeb M
UMR 6560, Laboratoire de Biochimie-Ingénierie des Protéines, Marseille, France.
Toxicon. 1998 Nov;36(11):1609-11. doi: 10.1016/s0041-0101(98)00153-6.
Maurotoxin, a toxin from the venom of the Tunisian chactoid scorpion Scorpio maurus, has been purified to homogeneity by gel filtration/reversed-phase HPLC, and characterized. It is a basic and C-terminal amidated 34-residue polypeptide cross-linked by four disulfide bridges. From Edman sequencing results, only six different pairings between the first six half-cystines were retained whereas a disulfide bridge was predicted between the two half-cystines in positions 31 and 34. Modelling based on the structure of charybdotoxin favored two different pairings, one of which possessed two disulfides in common with the general motif of scorpion toxins. The solid-phase technique was used to obtain synthetic maurotoxin, sMTX. The half-cystine pairings of sMTX were determined by enzymatic cleavage and were found to be Cys3 Cys24, Cys9-Cys29, Cys13-Cys19, and Cys31-34, in agreement with experimental data obtained with natural maurotoxin. Both natural and synthetic maurotoxins were lethal to mice following intracerebroventricular injection (LD50, 80 ng/mouse). They blocked the Kv1.1, Kv1.2, and Kv1.3 channels expressed in Xenopus oocytes with almost identical half-effects (IC50) in the range of 40, 0.8 and 150 nM, respectively. They also competed with 125I-apamin (SKca channel blocker) and 125I-kaliotoxin (Kv channel blocker) for binding to rat brain synaptosomes with IC50 of about 5 and 0.03 nM. As the natural and synthetic maurotoxins exhibit indistinguishable physicochemical and pharmacological properties, they are likely to adopt the same half-cystine pairing pattern which is unique among known scorpion toxins. However, this disulfide organization is different from those reported for Pandinus imperator and Heterometrus spinnifer toxins 1 (Pi1 and HsTx1), two novel four-disulfide bridged K+ channel-acting scorpion toxin sharing about 50-70% sequence identity with maurotoxin.
毛罗毒素是一种来自突尼斯钳蝎毒液的毒素,已通过凝胶过滤/反相高效液相色谱法纯化至同质,并进行了表征。它是一种碱性且C端酰胺化的34个残基的多肽,由四个二硫键交联。根据埃德曼测序结果,仅保留了前六个半胱氨酸之间六种不同的配对方式,而预测在第31和34位的两个半胱氨酸之间存在一个二硫键。基于蝎毒素结构的建模支持两种不同的配对方式,其中一种与蝎毒素的一般基序有两个共同的二硫键。采用固相技术获得了合成毛罗毒素(sMTX)。通过酶切确定了sMTX的半胱氨酸配对方式,发现为Cys3-Cys24、Cys9-Cys29、Cys13-Cys19和Cys31-34,这与天然毛罗毒素获得的实验数据一致。天然和合成毛罗毒素经脑室内注射后对小鼠均具有致死性(LD50,80 ng/小鼠)。它们阻断非洲爪蟾卵母细胞中表达的Kv1.1、Kv1.2和Kv1.3通道,其半数效应(IC50)几乎相同,分别在40、0.8和150 nM范围内。它们还与125I-蜂毒明肽(SKca通道阻滞剂)和125I-钾离子毒素(Kv通道阻滞剂)竞争与大鼠脑突触体的结合,IC50约为5和0.03 nM。由于天然和合成毛罗毒素表现出难以区分的物理化学和药理学特性,它们可能采用相同的半胱氨酸配对模式,这在已知的蝎毒素中是独特的。然而,这种二硫键结构与帝王蝎和棘刺异蝎毒素1(Pi1和HsTx1)报道的不同,这两种新型的四二硫键桥接的K+通道作用蝎毒素与毛罗毒素具有约50-70%的序列同一性。