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

M. R. V. Diniz; R.David G. Theakston; J. M. Crampton; M. D. Cordeiro; A. M. C. Pimenta; M. E. De Lima; C. R. Diniz

doi:10.1016/j.pep.2006.06.012

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

M. R. V. Diniz, R.David G. Theakston, J. M. Crampton, M. D. Cordeiro, A. M. C. Pimenta, M. E. De Lima, C. R. Diniz

Tropical Disease Biology

Liverpool School of Tropical Medicine

Research output: Contribution to journal › Article › peer-review

19 Citations (Scopus)

Abstract

Tx1 from the venom of the Brazilian spider, Phoneutria nigriventer, is a lethal neurotoxic polypeptide of M, 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. (c) 2006 Elsevier Inc. All rights reserved.

Original language	English
Pages (from-to)	18-24
Number of pages	7
Journal	Protein Expression and Purification
Volume	50
Issue number	1
DOIs	https://doi.org/10.1016/j.pep.2006.06.012
Publication status	Published - 1 Nov 2006

Keywords

Escherichia coli
Phoneutria nigriventer
Sodium channel
Spider toxin
Thioredoxin

Access to Document

10.1016/j.pep.2006.06.012

Cite this

Diniz, M. R. V., Theakston, R. D. G., Crampton, J. M., Cordeiro, M. D., Pimenta, A. M. C., De Lima, M. E., & Diniz, C. R. (2006). Functional expression and purification of recombinant Tx1, a sodium channel blocker neurotoxin from the venom of the Brazilian "armed" spider, Phoneutria nigriventer. Protein Expression and Purification, 50(1), 18-24. https://doi.org/10.1016/j.pep.2006.06.012

@article{0a40ce91fa95462dba8b33337aa26e5b,

title = "Functional expression and purification of recombinant Tx1, a sodium channel blocker neurotoxin from the venom of the Brazilian {"}armed{"} spider, Phoneutria nigriventer",

abstract = "Tx1 from the venom of the Brazilian spider, Phoneutria nigriventer, is a lethal neurotoxic polypeptide of M, 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. (c) 2006 Elsevier Inc. All rights reserved.",

keywords = "Escherichia coli, Phoneutria nigriventer, Sodium channel, Spider toxin, Thioredoxin",

author = "Diniz, \{M. R. V.\} and Theakston, \{R.David G.\} and Crampton, \{J. M.\} and Cordeiro, \{M. D.\} and Pimenta, \{A. M. C.\} and \{De Lima\}, \{M. E.\} and Diniz, \{C. R.\}",

year = "2006",

month = nov,

day = "1",

doi = "10.1016/j.pep.2006.06.012",

language = "English",

volume = "50",

pages = "18--24",

journal = "Protein Expression and Purification",

issn = "1046-5928",

publisher = "Academic Press Inc.",

number = "1",

}

Diniz, MRV, Theakston, RDG, Crampton, JM, Cordeiro, MD, Pimenta, AMC, De Lima, ME & Diniz, CR 2006, 'Functional expression and purification of recombinant Tx1, a sodium channel blocker neurotoxin from the venom of the Brazilian "armed" spider, Phoneutria nigriventer', Protein Expression and Purification, vol. 50, no. 1, pp. 18-24. https://doi.org/10.1016/j.pep.2006.06.012

Functional expression and purification of recombinant Tx1, a sodium channel blocker neurotoxin from the venom of the Brazilian "armed" spider, Phoneutria nigriventer. / Diniz, M. R. V.; Theakston, R.David G.; Crampton, J. M. et al.
In: Protein Expression and Purification, Vol. 50, No. 1, 01.11.2006, p. 18-24.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Functional expression and purification of recombinant Tx1, a sodium channel blocker neurotoxin from the venom of the Brazilian "armed" spider, Phoneutria nigriventer

AU - Diniz, M. R. V.

AU - Theakston, R.David G.

AU - Crampton, J. M.

AU - Cordeiro, M. D.

AU - Pimenta, A. M. C.

AU - De Lima, M. E.

AU - Diniz, C. R.

PY - 2006/11/1

Y1 - 2006/11/1

N2 - Tx1 from the venom of the Brazilian spider, Phoneutria nigriventer, is a lethal neurotoxic polypeptide of M, 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. (c) 2006 Elsevier Inc. All rights reserved.

AB - Tx1 from the venom of the Brazilian spider, Phoneutria nigriventer, is a lethal neurotoxic polypeptide of M, 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. (c) 2006 Elsevier Inc. All rights reserved.

KW - Escherichia coli

KW - Phoneutria nigriventer

KW - Sodium channel

KW - Spider toxin

KW - Thioredoxin

U2 - 10.1016/j.pep.2006.06.012

DO - 10.1016/j.pep.2006.06.012

M3 - Article

SN - 1046-5928

VL - 50

SP - 18

EP - 24

JO - Protein Expression and Purification

JF - Protein Expression and Purification

IS - 1

ER -

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

Abstract

Keywords

Access to Document

Fingerprint

Cite this