Impact of Q139R substitution of MEB4-Cry2Aa toxin on its stability, accessibility and toxicity against Ephestia kuehniella
The Bacillus thuringiensis subsp. kurstaki strain MEB4 was previously found to be highly toxic to Ephestia kuehniella. SDS-PAGE analysis of the recombinant strain DH5α (pBS-cry2Aa-MEB4) showed that Cry2Aa-MEB4 delta-endotoxins were forming inclusion bodies, and were 2.75 fold more toxic towards E. k...
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| Veröffentlicht in: | International journal of biological macromolecules 2015-11, Vol.81, p.701-709 |
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| creator | Nouha, Abdelmalek Sameh, Sellami Fakher, Frikha Slim, Tounsi Souad, Rouis |
| description | The Bacillus thuringiensis subsp. kurstaki strain MEB4 was previously found to be highly toxic to Ephestia kuehniella. SDS-PAGE analysis of the recombinant strain DH5α (pBS-cry2Aa-MEB4) showed that Cry2Aa-MEB4 delta-endotoxins were forming inclusion bodies, and were 2.75 fold more toxic towards E. kuehniella than those of Cry2Aa-BNS3. Besides to the 65kDa active toxin, proteolysis activation of Cry2Aa-BNS3 protein with E. kuehniella midgut juice generated an extra proteolysis form of 49kDa, which was the result of another chymotrypsin cleavage located in Leu144. The amino acid sequences alignment of Cry2Aa-MEB4 and Cry2Aa-BNS3 showed that among the different 15 amino acids, the Q139R substitution was found to be interesting. In fact, due to its presence within the loop α3-α4, the chymotrypsin-like protease was unable to access to its site in Cry2Aa-MEB4, resulting to the production of only the 65kDa form. The accessible surface and the stability studies of the structure model of the Cry2Aa-BNS3-49 form showed a lower hydrophobicity surface due to the omission of 144 amino acids from the N-terminal comparing with the active Cry2Aa-MEB4 protein. All these features caused the diminishing of Cry2Aa-BNS3 toxicity towards E. kuehniella. |
| doi_str_mv | 10.1016/j.ijbiomac.2015.08.058 |
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SDS-PAGE analysis of the recombinant strain DH5α (pBS-cry2Aa-MEB4) showed that Cry2Aa-MEB4 delta-endotoxins were forming inclusion bodies, and were 2.75 fold more toxic towards E. kuehniella than those of Cry2Aa-BNS3. Besides to the 65kDa active toxin, proteolysis activation of Cry2Aa-BNS3 protein with E. kuehniella midgut juice generated an extra proteolysis form of 49kDa, which was the result of another chymotrypsin cleavage located in Leu144. The amino acid sequences alignment of Cry2Aa-MEB4 and Cry2Aa-BNS3 showed that among the different 15 amino acids, the Q139R substitution was found to be interesting. In fact, due to its presence within the loop α3-α4, the chymotrypsin-like protease was unable to access to its site in Cry2Aa-MEB4, resulting to the production of only the 65kDa form. The accessible surface and the stability studies of the structure model of the Cry2Aa-BNS3-49 form showed a lower hydrophobicity surface due to the omission of 144 amino acids from the N-terminal comparing with the active Cry2Aa-MEB4 protein. All these features caused the diminishing of Cry2Aa-BNS3 toxicity towards E. kuehniella.</description><identifier>ISSN: 0141-8130</identifier><identifier>EISSN: 1879-0003</identifier><identifier>DOI: 10.1016/j.ijbiomac.2015.08.058</identifier><identifier>PMID: 26321422</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Amino Acid Sequence ; Amino Acid Substitution ; Animals ; Antibiosis ; Bacillus thuringiensis - genetics ; Bacillus thuringiensis - metabolism ; Codon ; Cry2Aa ; Endotoxins - chemistry ; Endotoxins - genetics ; Endotoxins - toxicity ; Gene Expression ; Hydrophobic and Hydrophilic Interactions ; Hydrophobicity surface ; Insect Control ; Models, Molecular ; Molecular Sequence Data ; Moths - microbiology ; Mutation ; Protein Conformation ; Protein Stability ; Proteolysis ; Recombinant Proteins - chemistry ; Recombinant Proteins - genetics ; Recombinant Proteins - toxicity ; Sequence Alignment ; Thermostability</subject><ispartof>International journal of biological macromolecules, 2015-11, Vol.81, p.701-709</ispartof><rights>2015 Elsevier B.V.</rights><rights>Copyright © 2015 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c368t-1184070ae1d7144dcf5030afb69ba1fc76fce4fb7ef73fad10def4275ba4733c3</citedby><cites>FETCH-LOGICAL-c368t-1184070ae1d7144dcf5030afb69ba1fc76fce4fb7ef73fad10def4275ba4733c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0141813015006054$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26321422$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Nouha, Abdelmalek</creatorcontrib><creatorcontrib>Sameh, Sellami</creatorcontrib><creatorcontrib>Fakher, Frikha</creatorcontrib><creatorcontrib>Slim, Tounsi</creatorcontrib><creatorcontrib>Souad, Rouis</creatorcontrib><title>Impact of Q139R substitution of MEB4-Cry2Aa toxin on its stability, accessibility and toxicity against Ephestia kuehniella</title><title>International journal of biological macromolecules</title><addtitle>Int J Biol Macromol</addtitle><description>The Bacillus thuringiensis subsp. kurstaki strain MEB4 was previously found to be highly toxic to Ephestia kuehniella. SDS-PAGE analysis of the recombinant strain DH5α (pBS-cry2Aa-MEB4) showed that Cry2Aa-MEB4 delta-endotoxins were forming inclusion bodies, and were 2.75 fold more toxic towards E. kuehniella than those of Cry2Aa-BNS3. Besides to the 65kDa active toxin, proteolysis activation of Cry2Aa-BNS3 protein with E. kuehniella midgut juice generated an extra proteolysis form of 49kDa, which was the result of another chymotrypsin cleavage located in Leu144. The amino acid sequences alignment of Cry2Aa-MEB4 and Cry2Aa-BNS3 showed that among the different 15 amino acids, the Q139R substitution was found to be interesting. In fact, due to its presence within the loop α3-α4, the chymotrypsin-like protease was unable to access to its site in Cry2Aa-MEB4, resulting to the production of only the 65kDa form. The accessible surface and the stability studies of the structure model of the Cry2Aa-BNS3-49 form showed a lower hydrophobicity surface due to the omission of 144 amino acids from the N-terminal comparing with the active Cry2Aa-MEB4 protein. All these features caused the diminishing of Cry2Aa-BNS3 toxicity towards E. kuehniella.</description><subject>Amino Acid Sequence</subject><subject>Amino Acid Substitution</subject><subject>Animals</subject><subject>Antibiosis</subject><subject>Bacillus thuringiensis - genetics</subject><subject>Bacillus thuringiensis - metabolism</subject><subject>Codon</subject><subject>Cry2Aa</subject><subject>Endotoxins - chemistry</subject><subject>Endotoxins - genetics</subject><subject>Endotoxins - toxicity</subject><subject>Gene Expression</subject><subject>Hydrophobic and Hydrophilic Interactions</subject><subject>Hydrophobicity surface</subject><subject>Insect Control</subject><subject>Models, Molecular</subject><subject>Molecular Sequence Data</subject><subject>Moths - microbiology</subject><subject>Mutation</subject><subject>Protein Conformation</subject><subject>Protein Stability</subject><subject>Proteolysis</subject><subject>Recombinant Proteins - chemistry</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - toxicity</subject><subject>Sequence Alignment</subject><subject>Thermostability</subject><issn>0141-8130</issn><issn>1879-0003</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1v1DAQhi0EotuWv1D52AMJntiJk1vb1UIrFSFQOVuOM25nu0m2sYNYfj3ebsuVy4zm1TtfD2NnIHIQUH1a57RuaeytywsBZS7qXJT1G7aAWjeZEEK-ZQsBCrIapDhixyGsk1qVUL9nR0UlC1BFsWB_bvqtdZGPnn8H2fzgYW5DpDhHGoe9-nV1pbLltCsuLY_jb0riwCkGHqJtaUNx95Fb5zAEOpTcDt2z0z0X95aGEPlq-4BpruWPMz4MhJuNPWXvvN0E_PCST9jPz6u75XV2--3LzfLyNnOyqmMGUCuhhUXoNCjVOV8KKaxvq6a14J2uvEPlW41eS287EB16VeiytUpL6eQJOz_M3U7j05yuMD0Ft79gwHEOBnQJqkmxSdbqYHXTGMKE3mwn6u20MyDMnrtZm1fuZs_diNok7qnx7GXH3PbY_Wt7BZ0MFwcDpk9_EU4mOMLBYUcTumi6kf634y_3t5jS</recordid><startdate>201511</startdate><enddate>201511</enddate><creator>Nouha, Abdelmalek</creator><creator>Sameh, Sellami</creator><creator>Fakher, Frikha</creator><creator>Slim, Tounsi</creator><creator>Souad, Rouis</creator><general>Elsevier B.V</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>7X8</scope></search><sort><creationdate>201511</creationdate><title>Impact of Q139R substitution of MEB4-Cry2Aa toxin on its stability, accessibility and toxicity against Ephestia kuehniella</title><author>Nouha, Abdelmalek ; Sameh, Sellami ; Fakher, Frikha ; Slim, Tounsi ; Souad, Rouis</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c368t-1184070ae1d7144dcf5030afb69ba1fc76fce4fb7ef73fad10def4275ba4733c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Amino Acid Sequence</topic><topic>Amino Acid Substitution</topic><topic>Animals</topic><topic>Antibiosis</topic><topic>Bacillus thuringiensis - genetics</topic><topic>Bacillus thuringiensis - metabolism</topic><topic>Codon</topic><topic>Cry2Aa</topic><topic>Endotoxins - chemistry</topic><topic>Endotoxins - genetics</topic><topic>Endotoxins - toxicity</topic><topic>Gene Expression</topic><topic>Hydrophobic and Hydrophilic Interactions</topic><topic>Hydrophobicity surface</topic><topic>Insect Control</topic><topic>Models, Molecular</topic><topic>Molecular Sequence Data</topic><topic>Moths - microbiology</topic><topic>Mutation</topic><topic>Protein Conformation</topic><topic>Protein Stability</topic><topic>Proteolysis</topic><topic>Recombinant Proteins - chemistry</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - toxicity</topic><topic>Sequence Alignment</topic><topic>Thermostability</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nouha, Abdelmalek</creatorcontrib><creatorcontrib>Sameh, Sellami</creatorcontrib><creatorcontrib>Fakher, Frikha</creatorcontrib><creatorcontrib>Slim, Tounsi</creatorcontrib><creatorcontrib>Souad, Rouis</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>International journal of biological macromolecules</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nouha, Abdelmalek</au><au>Sameh, Sellami</au><au>Fakher, Frikha</au><au>Slim, Tounsi</au><au>Souad, Rouis</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Impact of Q139R substitution of MEB4-Cry2Aa toxin on its stability, accessibility and toxicity against Ephestia kuehniella</atitle><jtitle>International journal of biological macromolecules</jtitle><addtitle>Int J Biol Macromol</addtitle><date>2015-11</date><risdate>2015</risdate><volume>81</volume><spage>701</spage><epage>709</epage><pages>701-709</pages><issn>0141-8130</issn><eissn>1879-0003</eissn><abstract>The Bacillus thuringiensis subsp. kurstaki strain MEB4 was previously found to be highly toxic to Ephestia kuehniella. SDS-PAGE analysis of the recombinant strain DH5α (pBS-cry2Aa-MEB4) showed that Cry2Aa-MEB4 delta-endotoxins were forming inclusion bodies, and were 2.75 fold more toxic towards E. kuehniella than those of Cry2Aa-BNS3. Besides to the 65kDa active toxin, proteolysis activation of Cry2Aa-BNS3 protein with E. kuehniella midgut juice generated an extra proteolysis form of 49kDa, which was the result of another chymotrypsin cleavage located in Leu144. The amino acid sequences alignment of Cry2Aa-MEB4 and Cry2Aa-BNS3 showed that among the different 15 amino acids, the Q139R substitution was found to be interesting. In fact, due to its presence within the loop α3-α4, the chymotrypsin-like protease was unable to access to its site in Cry2Aa-MEB4, resulting to the production of only the 65kDa form. The accessible surface and the stability studies of the structure model of the Cry2Aa-BNS3-49 form showed a lower hydrophobicity surface due to the omission of 144 amino acids from the N-terminal comparing with the active Cry2Aa-MEB4 protein. All these features caused the diminishing of Cry2Aa-BNS3 toxicity towards E. kuehniella.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>26321422</pmid><doi>10.1016/j.ijbiomac.2015.08.058</doi><tpages>9</tpages></addata></record> |
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| subjects | Amino Acid Sequence Amino Acid Substitution Animals Antibiosis Bacillus thuringiensis - genetics Bacillus thuringiensis - metabolism Codon Cry2Aa Endotoxins - chemistry Endotoxins - genetics Endotoxins - toxicity Gene Expression Hydrophobic and Hydrophilic Interactions Hydrophobicity surface Insect Control Models, Molecular Molecular Sequence Data Moths - microbiology Mutation Protein Conformation Protein Stability Proteolysis Recombinant Proteins - chemistry Recombinant Proteins - genetics Recombinant Proteins - toxicity Sequence Alignment Thermostability |
| title | Impact of Q139R substitution of MEB4-Cry2Aa toxin on its stability, accessibility and toxicity against Ephestia kuehniella |
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