The purification and amino acid sequence of the lethal neurotoxin Tx1 from the venom of the Brazilian ‘armed’ spider Phoneutria nigriventer

FEBS Letters

do Nascimento Cordeiro, Marta¹  Junor, Leonides Rezende¹  Diniz, Carlos Ribeiro¹  Fischer, Stefan⁴  Reimann, Frank⁴  Kelly, Patrick²  Richardson, Michael³  Oliveira, Eduardo B.⁵ 

[1] Fundacão Ezequiel Dias, Rua Conde Pereira Carneiro, 30.550 Belo Horizonte (M. G.), Brazil;Applied Biosystems Ltd., Warrington, Cheshire WA3 7PB, England;Department of Biological Sciences, University of Durham, Durham DH1 3LE, England;Knauer Wiss. Gerätebau, Hegauer Weg 38, D-1000 Berlin, FRG;Departamento de Bioquimica, Faculdade de Medicina, Ribeirão Preto, São Paulo, Brazil

关键词: Spider venom;    Amino acid sequence;    Polypeptide neurotoxin;    Phoneutria nigriventer;   

DOI  :  10.1016/0014-5793(90)81386-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A lethal neurotoxic polypeptide of M_r 8kDa 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.

【 授权许可】

Unknown