Evidence for intermolecular interaction as a necessary step for pore-formation activity and toxicity of Bacillus thuringiensis Cry1Ab toxin
Based on the observation of large conductance states formed by Bacillus thuringiensis Cry toxins in synthetic planar lipid bilayers and the estimation of a pore size of 10–20 Å, it has been proposed that the pore could be formed by an oligomer containing four to six Cry toxin monomers. However, ther...
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| Veröffentlicht in: | FEMS microbiology letters 2000-10, Vol.191 (2), p.221-225 |
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| creator | Soberón, Mario Pérez, Rigoberto V. Nuñez-Valdéz, Marı́a E. Lorence, Argelia Gómez, Isabel Sánchez, Jorge Bravo, Alejandra |
| description | Based on the observation of large conductance states formed by
Bacillus thuringiensis Cry toxins in synthetic planar lipid bilayers and the estimation of a pore size of 10–20 Å, it has been proposed that the pore could be formed by an oligomer containing four to six Cry toxin monomers. However, there is a lack of information regarding the insertion of Cry toxins into the membrane and oligomer formation. Here we provide direct evidence showing that the intermolecular interaction between Cry1Ab toxin monomers is a necessary step for pore formation and toxicity. Two Cry1Ab mutant proteins affected in different steps of their mode of action (F371A in receptor binding and H168F in pore formation) were affected in toxicity against
Manduca sexta larvae. Binding analysis showed that F371A protein bound more efficiently to
M. sexta brush border membrane vesicles when mixed with H168F in a one to one ratio. These mutant proteins also recovered pore-formation activity, measured with a fluorescent dye with isolated brush border membrane vesicles, and toxicity against
M. sexta larvae when mixed, showing that monomers affected in different steps of their mode of action can form functional hetero-oligomers. |
| doi_str_mv | 10.1016/S0378-1097(00)00394-3 |
| format | Article |
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Bacillus thuringiensis Cry toxins in synthetic planar lipid bilayers and the estimation of a pore size of 10–20 Å, it has been proposed that the pore could be formed by an oligomer containing four to six Cry toxin monomers. However, there is a lack of information regarding the insertion of Cry toxins into the membrane and oligomer formation. Here we provide direct evidence showing that the intermolecular interaction between Cry1Ab toxin monomers is a necessary step for pore formation and toxicity. Two Cry1Ab mutant proteins affected in different steps of their mode of action (F371A in receptor binding and H168F in pore formation) were affected in toxicity against
Manduca sexta larvae. Binding analysis showed that F371A protein bound more efficiently to
M. sexta brush border membrane vesicles when mixed with H168F in a one to one ratio. These mutant proteins also recovered pore-formation activity, measured with a fluorescent dye with isolated brush border membrane vesicles, and toxicity against
M. sexta larvae when mixed, showing that monomers affected in different steps of their mode of action can form functional hetero-oligomers.</description><identifier>ISSN: 0378-1097</identifier><identifier>EISSN: 1574-6968</identifier><identifier>DOI: 10.1016/S0378-1097(00)00394-3</identifier><identifier>PMID: 11024267</identifier><identifier>CODEN: FMLED7</identifier><language>eng</language><publisher>Oxford: Elsevier B.V</publisher><subject>Animals ; Bacillus thuringiensis ; Bacillus thuringiensis - genetics ; Bacillus thuringiensis - metabolism ; Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; Bacterial Proteins - toxicity ; Bacterial Toxins ; Bacteriology ; Biological and medical sciences ; brush border membrane vesicles ; Cell Membrane Structures - metabolism ; Cry1Ab protein ; Endotoxins - genetics ; Endotoxins - metabolism ; Endotoxins - toxicity ; fluorescent dyes ; Fundamental and applied biological sciences. Psychology ; Hemolysin Proteins ; Ion Channels - physiology ; Larva - drug effects ; larvae ; lipid bilayers ; Manduca - drug effects ; Manduca - growth & development ; Manduca - metabolism ; Manduca sexta ; mechanism of action ; Membrane Potentials - drug effects ; Microbiology ; Microvilli ; mutants ; Mutation ; Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains ; Pore formation ; porosity ; proteins ; toxicity ; toxins ; δ-Endotoxin oligomerization</subject><ispartof>FEMS microbiology letters, 2000-10, Vol.191 (2), p.221-225</ispartof><rights>2000 Federation of European Microbiological Societies</rights><rights>2001 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c444t-ee95f0f316d8e3b8347beeda06ba4b1531be3c471d39c4d915a30acb5864bfd43</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=993824$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11024267$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Soberón, Mario</creatorcontrib><creatorcontrib>Pérez, Rigoberto V.</creatorcontrib><creatorcontrib>Nuñez-Valdéz, Marı́a E.</creatorcontrib><creatorcontrib>Lorence, Argelia</creatorcontrib><creatorcontrib>Gómez, Isabel</creatorcontrib><creatorcontrib>Sánchez, Jorge</creatorcontrib><creatorcontrib>Bravo, Alejandra</creatorcontrib><title>Evidence for intermolecular interaction as a necessary step for pore-formation activity and toxicity of Bacillus thuringiensis Cry1Ab toxin</title><title>FEMS microbiology letters</title><addtitle>FEMS Microbiol Lett</addtitle><description>Based on the observation of large conductance states formed by
Bacillus thuringiensis Cry toxins in synthetic planar lipid bilayers and the estimation of a pore size of 10–20 Å, it has been proposed that the pore could be formed by an oligomer containing four to six Cry toxin monomers. However, there is a lack of information regarding the insertion of Cry toxins into the membrane and oligomer formation. Here we provide direct evidence showing that the intermolecular interaction between Cry1Ab toxin monomers is a necessary step for pore formation and toxicity. Two Cry1Ab mutant proteins affected in different steps of their mode of action (F371A in receptor binding and H168F in pore formation) were affected in toxicity against
Manduca sexta larvae. Binding analysis showed that F371A protein bound more efficiently to
M. sexta brush border membrane vesicles when mixed with H168F in a one to one ratio. These mutant proteins also recovered pore-formation activity, measured with a fluorescent dye with isolated brush border membrane vesicles, and toxicity against
M. sexta larvae when mixed, showing that monomers affected in different steps of their mode of action can form functional hetero-oligomers.</description><subject>Animals</subject><subject>Bacillus thuringiensis</subject><subject>Bacillus thuringiensis - genetics</subject><subject>Bacillus thuringiensis - metabolism</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - metabolism</subject><subject>Bacterial Proteins - toxicity</subject><subject>Bacterial Toxins</subject><subject>Bacteriology</subject><subject>Biological and medical sciences</subject><subject>brush border membrane vesicles</subject><subject>Cell Membrane Structures - metabolism</subject><subject>Cry1Ab protein</subject><subject>Endotoxins - genetics</subject><subject>Endotoxins - metabolism</subject><subject>Endotoxins - toxicity</subject><subject>fluorescent dyes</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Hemolysin Proteins</subject><subject>Ion Channels - physiology</subject><subject>Larva - drug effects</subject><subject>larvae</subject><subject>lipid bilayers</subject><subject>Manduca - drug effects</subject><subject>Manduca - growth & development</subject><subject>Manduca - metabolism</subject><subject>Manduca sexta</subject><subject>mechanism of action</subject><subject>Membrane Potentials - drug effects</subject><subject>Microbiology</subject><subject>Microvilli</subject><subject>mutants</subject><subject>Mutation</subject><subject>Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains</subject><subject>Pore formation</subject><subject>porosity</subject><subject>proteins</subject><subject>toxicity</subject><subject>toxins</subject><subject>δ-Endotoxin oligomerization</subject><issn>0378-1097</issn><issn>1574-6968</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0c1u1DAQB_AIgei28AiAJaQKDoFx7DjxCZVV-ZAqcSg9W449KUaJvdjJin0GXhpnsypHTral39jj-RfFCwrvKFDx_hZY05YUZPMG4C0Ak7xkj4oNrRteCinax8XmgZwV5yn9BABegXhanFEKFa9Esyn-XO-dRW-Q9CES5yeMYxjQzIM-HbWZXPBEJ6KJR4Mp6XggacLdsWQXIpZ5M-qVZb1304Fob8kUfjuzHEJPPmrjhmFOZPoxR-fvHfrkEtnGA73qjtI_K570ekj4_LReFHefrr9vv5Q33z5_3V7dlIZzPpWIsu6hZ1TYFlnXMt50iFaD6DTvaM1oh8zwhlomDbeS1pqBNl3dCt71lrOL4nK9dxfDrxnTpEaXDA6D9hjmpGgjWmgamWG9QhNDShF7tYtuzN9XFNSSgjqmoJYRKwB1TEGxXPfy9MDcjWj_VZ3GnsHrE9DJ6KGP2huXHpyUrK2WPl-tqtdB6fuYxd1tlZ_NQUpRs0V8WAXmce0dRpWMW9K0LqKZlA3uP63-BdRUsFE</recordid><startdate>20001015</startdate><enddate>20001015</enddate><creator>Soberón, Mario</creator><creator>Pérez, Rigoberto V.</creator><creator>Nuñez-Valdéz, Marı́a E.</creator><creator>Lorence, Argelia</creator><creator>Gómez, Isabel</creator><creator>Sánchez, Jorge</creator><creator>Bravo, Alejandra</creator><general>Elsevier B.V</general><general>Blackwell</general><scope>FBQ</scope><scope>IQODW</scope><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>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope></search><sort><creationdate>20001015</creationdate><title>Evidence for intermolecular interaction as a necessary step for pore-formation activity and toxicity of Bacillus thuringiensis Cry1Ab toxin</title><author>Soberón, Mario ; Pérez, Rigoberto V. ; Nuñez-Valdéz, Marı́a E. ; Lorence, Argelia ; Gómez, Isabel ; Sánchez, Jorge ; Bravo, Alejandra</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c444t-ee95f0f316d8e3b8347beeda06ba4b1531be3c471d39c4d915a30acb5864bfd43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Animals</topic><topic>Bacillus thuringiensis</topic><topic>Bacillus thuringiensis - genetics</topic><topic>Bacillus thuringiensis - metabolism</topic><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Proteins - metabolism</topic><topic>Bacterial Proteins - toxicity</topic><topic>Bacterial Toxins</topic><topic>Bacteriology</topic><topic>Biological and medical sciences</topic><topic>brush border membrane vesicles</topic><topic>Cell Membrane Structures - metabolism</topic><topic>Cry1Ab protein</topic><topic>Endotoxins - genetics</topic><topic>Endotoxins - metabolism</topic><topic>Endotoxins - toxicity</topic><topic>fluorescent dyes</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Hemolysin Proteins</topic><topic>Ion Channels - physiology</topic><topic>Larva - drug effects</topic><topic>larvae</topic><topic>lipid bilayers</topic><topic>Manduca - drug effects</topic><topic>Manduca - growth & development</topic><topic>Manduca - metabolism</topic><topic>Manduca sexta</topic><topic>mechanism of action</topic><topic>Membrane Potentials - drug effects</topic><topic>Microbiology</topic><topic>Microvilli</topic><topic>mutants</topic><topic>Mutation</topic><topic>Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains</topic><topic>Pore formation</topic><topic>porosity</topic><topic>proteins</topic><topic>toxicity</topic><topic>toxins</topic><topic>δ-Endotoxin oligomerization</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Soberón, Mario</creatorcontrib><creatorcontrib>Pérez, Rigoberto V.</creatorcontrib><creatorcontrib>Nuñez-Valdéz, Marı́a E.</creatorcontrib><creatorcontrib>Lorence, Argelia</creatorcontrib><creatorcontrib>Gómez, Isabel</creatorcontrib><creatorcontrib>Sánchez, Jorge</creatorcontrib><creatorcontrib>Bravo, Alejandra</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><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>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>FEMS microbiology letters</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Soberón, Mario</au><au>Pérez, Rigoberto V.</au><au>Nuñez-Valdéz, Marı́a E.</au><au>Lorence, Argelia</au><au>Gómez, Isabel</au><au>Sánchez, Jorge</au><au>Bravo, Alejandra</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Evidence for intermolecular interaction as a necessary step for pore-formation activity and toxicity of Bacillus thuringiensis Cry1Ab toxin</atitle><jtitle>FEMS microbiology letters</jtitle><addtitle>FEMS Microbiol Lett</addtitle><date>2000-10-15</date><risdate>2000</risdate><volume>191</volume><issue>2</issue><spage>221</spage><epage>225</epage><pages>221-225</pages><issn>0378-1097</issn><eissn>1574-6968</eissn><coden>FMLED7</coden><abstract>Based on the observation of large conductance states formed by
Bacillus thuringiensis Cry toxins in synthetic planar lipid bilayers and the estimation of a pore size of 10–20 Å, it has been proposed that the pore could be formed by an oligomer containing four to six Cry toxin monomers. However, there is a lack of information regarding the insertion of Cry toxins into the membrane and oligomer formation. Here we provide direct evidence showing that the intermolecular interaction between Cry1Ab toxin monomers is a necessary step for pore formation and toxicity. Two Cry1Ab mutant proteins affected in different steps of their mode of action (F371A in receptor binding and H168F in pore formation) were affected in toxicity against
Manduca sexta larvae. Binding analysis showed that F371A protein bound more efficiently to
M. sexta brush border membrane vesicles when mixed with H168F in a one to one ratio. These mutant proteins also recovered pore-formation activity, measured with a fluorescent dye with isolated brush border membrane vesicles, and toxicity against
M. sexta larvae when mixed, showing that monomers affected in different steps of their mode of action can form functional hetero-oligomers.</abstract><cop>Oxford</cop><pub>Elsevier B.V</pub><pmid>11024267</pmid><doi>10.1016/S0378-1097(00)00394-3</doi><tpages>5</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0378-1097 |
| ispartof | FEMS microbiology letters, 2000-10, Vol.191 (2), p.221-225 |
| issn | 0378-1097 1574-6968 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_17680779 |
| source | MEDLINE; Access via Wiley Online Library; Oxford University Press Journals All Titles (1996-Current); Alma/SFX Local Collection |
| subjects | Animals Bacillus thuringiensis Bacillus thuringiensis - genetics Bacillus thuringiensis - metabolism Bacterial Proteins - genetics Bacterial Proteins - metabolism Bacterial Proteins - toxicity Bacterial Toxins Bacteriology Biological and medical sciences brush border membrane vesicles Cell Membrane Structures - metabolism Cry1Ab protein Endotoxins - genetics Endotoxins - metabolism Endotoxins - toxicity fluorescent dyes Fundamental and applied biological sciences. Psychology Hemolysin Proteins Ion Channels - physiology Larva - drug effects larvae lipid bilayers Manduca - drug effects Manduca - growth & development Manduca - metabolism Manduca sexta mechanism of action Membrane Potentials - drug effects Microbiology Microvilli mutants Mutation Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains Pore formation porosity proteins toxicity toxins δ-Endotoxin oligomerization |
| title | Evidence for intermolecular interaction as a necessary step for pore-formation activity and toxicity of Bacillus thuringiensis Cry1Ab toxin |
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