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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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| Published in: | Protein expression and purification 2006-11, Vol.50 (1), p.18-24 |
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| Main Authors: | , , , , , , |
| Format: | Article |
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| cited_by | cdi_FETCH-LOGICAL-c382t-536286b4a2333ed39de575fdd900d0b0d22f843a66b75950095f5df28f1adca63 |
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| cites | cdi_FETCH-LOGICAL-c382t-536286b4a2333ed39de575fdd900d0b0d22f843a66b75950095f5df28f1adca63 |
| container_end_page | 24 |
| container_issue | 1 |
| container_start_page | 18 |
| container_title | Protein expression and purification |
| container_volume | 50 |
| creator | 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. |
| description | 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. |
| doi_str_mv | 10.1016/j.pep.2006.06.012 |
| format | article |
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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.</description><identifier>ISSN: 1046-5928</identifier><identifier>EISSN: 1096-0279</identifier><identifier>DOI: 10.1016/j.pep.2006.06.012</identifier><identifier>PMID: 16908187</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>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</subject><ispartof>Protein expression and purification, 2006-11, Vol.50 (1), p.18-24</ispartof><rights>2006 Elsevier Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c382t-536286b4a2333ed39de575fdd900d0b0d22f843a66b75950095f5df28f1adca63</citedby><cites>FETCH-LOGICAL-c382t-536286b4a2333ed39de575fdd900d0b0d22f843a66b75950095f5df28f1adca63</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16908187$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Diniz, Marcelo R.V.</creatorcontrib><creatorcontrib>Theakston, R. David G.</creatorcontrib><creatorcontrib>Crampton, Julian M.</creatorcontrib><creatorcontrib>Nascimento Cordeiro, Marta do</creatorcontrib><creatorcontrib>Pimenta, Adriano M.C.</creatorcontrib><creatorcontrib>De Lima, Maria Elena</creatorcontrib><creatorcontrib>Diniz, Carlos R.</creatorcontrib><title>Functional expression and purification of recombinant Tx1, a sodium channel blocker neurotoxin from the venom of the Brazilian “armed” spider, Phoneutria nigriventer</title><title>Protein expression and purification</title><addtitle>Protein Expr Purif</addtitle><description>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.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Araneae</subject><subject>Binding, Competitive - drug effects</subject><subject>Brazil</subject><subject>CHO Cells</subject><subject>Cloning, Molecular</subject><subject>Cricetinae</subject><subject>Escherichia coli</subject><subject>Injections, Intraventricular</subject><subject>Mice</subject><subject>Molecular Sequence Data</subject><subject>Neuropeptides - biosynthesis</subject><subject>Neuropeptides - isolation & purification</subject><subject>Neuropeptides - toxicity</subject><subject>Phoneutria nigriventer</subject><subject>Protein Binding</subject><subject>Rats</subject><subject>Rats, Wistar</subject><subject>Recombinant Proteins - biosynthesis</subject><subject>Recombinant Proteins - isolation & purification</subject><subject>Recombinant Proteins - toxicity</subject><subject>Sodium - metabolism</subject><subject>Sodium channel</subject><subject>Sodium Channel Blockers - isolation & purification</subject><subject>Sodium Channel Blockers - metabolism</subject><subject>Sodium Channel Blockers - toxicity</subject><subject>Sodium Channels - drug effects</subject><subject>Sodium Channels - metabolism</subject><subject>Species Specificity</subject><subject>Spider toxin</subject><subject>Spider Venoms - chemistry</subject><subject>Spider Venoms - genetics</subject><subject>Spiders - genetics</subject><subject>Structure-Activity Relationship</subject><subject>Thioredoxin</subject><issn>1046-5928</issn><issn>1096-0279</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><recordid>eNp9kcFu1DAQhiMEoqXwAFyQT5yaZeysvbE4QUULUiU4lLPl2GPWS2IHO6kWTn0QeABeq09SR7sSN6SRPCP__yfN_FX1ksKKAhVvdqsRxxUDEKulKHtUnVKQoga2kY-Xfi1qLll7Uj3LeQdAqQD-tDqhQkJL281p9fdyDmbyMeie4H5MmHMZiA6WjHPyzhu9_JLoSEITh84HHSZys6fnRJMcrZ8HYrY6BOxJ10fzHRMJOKc4xb0PxKU4kGmL5BZD6QpmGd4n_cv3Xgdyf_dbpwHt_d0fkkdvMZ2TL9tYCFPymgT_LflinTA9r5443Wd8cXzPqq-XH24uPtbXn68-Xby7rk3TsqnmjWCt6NaaNU2DtpEW-YY7ayWAhQ4sY65dN1qIbsMlB5DccetY66i2RovmrHp94I4p_pgxT2rw2WDf64BxzorKFkQL6yKkB6FJMeeETo3JDzr9VBTUko_aqZKPWvJRS1FWPK-O8LkrW_9zHAMpgrcHAZYVbz0mlY3HYND6cv9J2ej_g38A_aamdA</recordid><startdate>20061101</startdate><enddate>20061101</enddate><creator>Diniz, Marcelo R.V.</creator><creator>Theakston, R. David G.</creator><creator>Crampton, Julian M.</creator><creator>Nascimento Cordeiro, Marta do</creator><creator>Pimenta, Adriano M.C.</creator><creator>De Lima, Maria Elena</creator><creator>Diniz, Carlos R.</creator><general>Elsevier Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7SS</scope><scope>C1K</scope></search><sort><creationdate>20061101</creationdate><title>Functional expression and purification of recombinant Tx1, a sodium channel blocker neurotoxin from the venom of the Brazilian “armed” spider, Phoneutria nigriventer</title><author>Diniz, Marcelo R.V. ; Theakston, R. 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David G.</creatorcontrib><creatorcontrib>Crampton, Julian M.</creatorcontrib><creatorcontrib>Nascimento Cordeiro, Marta do</creatorcontrib><creatorcontrib>Pimenta, Adriano M.C.</creatorcontrib><creatorcontrib>De Lima, Maria Elena</creatorcontrib><creatorcontrib>Diniz, Carlos R.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Environmental Sciences and Pollution Management</collection><jtitle>Protein expression and purification</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Diniz, Marcelo R.V.</au><au>Theakston, R. David G.</au><au>Crampton, Julian M.</au><au>Nascimento Cordeiro, Marta do</au><au>Pimenta, Adriano M.C.</au><au>De Lima, Maria Elena</au><au>Diniz, Carlos R.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Functional expression and purification of recombinant Tx1, a sodium channel blocker neurotoxin from the venom of the Brazilian “armed” spider, Phoneutria nigriventer</atitle><jtitle>Protein expression and purification</jtitle><addtitle>Protein Expr Purif</addtitle><date>2006-11-01</date><risdate>2006</risdate><volume>50</volume><issue>1</issue><spage>18</spage><epage>24</epage><pages>18-24</pages><issn>1046-5928</issn><eissn>1096-0279</eissn><abstract>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
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| fulltext | fulltext |
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| source | ScienceDirect Journals |
| 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 |
| title | Functional expression and purification of recombinant Tx1, a sodium channel blocker neurotoxin from the venom of the Brazilian “armed” spider, Phoneutria nigriventer |
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