Functional expression and purification of recombinant Tx1, a sodium channel blocker neurotoxin from the venom of the Brazilian “armed” spider, Phoneutria nigriventer

Tx1 from the venom of the Brazilian spider, Phoneutria nigriventer, is a lethal neurotoxic polypeptide of M r 8600 Da with 14 cysteine residues. It is a novel sodium channel blocker which reversibly inhibits sodium currents in CHO cells expressing recombinant sodium (Nav1.2) channels. We cloned and...

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Bibliographic Details
Published in:Protein expression and purification 2006-11, Vol.50 (1), p.18-24
Main Authors: Diniz, Marcelo R.V., Theakston, R. David G., Crampton, Julian M., Nascimento Cordeiro, Marta do, Pimenta, Adriano M.C., De Lima, Maria Elena, Diniz, Carlos R.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Araneae
Binding, Competitive - drug effects
Brazil
CHO Cells
Cloning, Molecular
Cricetinae
Escherichia coli
Injections, Intraventricular
Mice
Molecular Sequence Data
Neuropeptides - biosynthesis
Neuropeptides - isolation & purification
Neuropeptides - toxicity
Phoneutria nigriventer
Protein Binding
Rats
Rats, Wistar
Recombinant Proteins - biosynthesis
Recombinant Proteins - isolation & purification
Recombinant Proteins - toxicity
Sodium - metabolism
Sodium channel
Sodium Channel Blockers - isolation & purification
Sodium Channel Blockers - metabolism
Sodium Channel Blockers - toxicity
Sodium Channels - drug effects
Sodium Channels - metabolism
Species Specificity
Spider toxin
Spider Venoms - chemistry
Spider Venoms - genetics
Spiders - genetics
Structure-Activity Relationship
Thioredoxin
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Description
Summary:Tx1 from the venom of the Brazilian spider, Phoneutria nigriventer, is a lethal neurotoxic polypeptide of M r 8600 Da with 14 cysteine residues. It is a novel sodium channel blocker which reversibly inhibits sodium currents in CHO cells expressing recombinant sodium (Nav1.2) channels. We cloned and expressed the Tx1 toxin as a thioredoxin fusion product in the cytoplasm of Escherichia coli. After semipurification by immobilized Ni–ion affinity chromatography, the recombinant Tx1 was purified by reverse phase chromatography and characterized. It displayed similar biochemical and pharmacological properties to the native toxin, and it should be useful for further investigation of structure-function relationship of Na channels.
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2006.06.012