Diniz C R, Cordeiro M do N, Junor L R, Kelly P, Fischer S, Reimann F, Oliveira E B, Richardson M
Fundacão Ezequiel Dias, Rua conde Pereira Carneiro, Belo Horizonte, Brazil.
FEBS Lett. 1990 Apr 24;263(2):251-3. doi: 10.1016/0014-5793(90)81386-3.
A lethal neurotoxic polypeptide of Mr 8 kDa was purified from the venom of the South American 'armed' or wandering spider Phoneutria nigriventer by centrifugation, gel filtration on Superose 12, and reverse phase FPLC on columns of Pharmacia PepRPC and ProRPC. The purified neurotoxin Tx1 had an LD50 of 0.05 mg/kg in mice following intracerebroventricular injection. The complete amino acid sequence of the neurotoxin was determined by automated Edman degradation of the native and S-carboxymethylated protein in pulsed liquid and dual phase sequencers, and by the manual DABITC/PITC double coupling method applied to fragments obtained after digestions with the S. aureus V8 protease and trypsin. The neurotoxin Tx1 consists of a single chain of 77 amino acid residues, which contains a high proportion of cysteine. The primary structure showed no homology to other identified spider toxins.
通过离心、在Superose 12上进行凝胶过滤以及在Pharmacia PepRPC和ProRPC柱上进行反相快速蛋白质液相色谱，从南美“武装”或游走蜘蛛黑腹捕鸟蛛的毒液中纯化出一种分子量为8 kDa的致死性神经毒性多肽。纯化后的神经毒素Tx1经脑室内注射，在小鼠中的半数致死剂量为0.05 mg/kg。通过在脉冲液相和双相测序仪中对天然和S-羧甲基化蛋白进行自动埃德曼降解，以及对金黄色葡萄球菌V8蛋白酶和胰蛋白酶消化后获得的片段应用手动DABITC/PITC双偶联法，确定了神经毒素的完整氨基酸序列。神经毒素Tx1由一条包含77个氨基酸残基的单链组成，其中半胱氨酸比例很高。其一级结构与其他已鉴定的蜘蛛毒素没有同源性。
