Two disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa δ-endotoxin increase stability with no effect on toxicity
9 p.
- Autores:
-
Wu, Sheng Jiun
Florez, Alvaro M.
Homoelle, Bradley J.
Dean, Donald H.
Alzate, Oscar
- Tipo de recurso:
- Article of journal
- Fecha de publicación:
- 2012
- Institución:
- Universidad de Santander
- Repositorio:
- Repositorio Universidad de Santander
- Idioma:
- eng
- OAI Identifier:
- oai:repositorio.udes.edu.co:001/3659
- Acceso en línea:
- https://repositorio.udes.edu.co/handle/001/3659
- Palabra clave:
- Disulfide Bonds
CD Spectra
Cry3Aa
Site Directed Mutagenesis
- Rights
- openAccess
- License
- Derechos Reservados - Universidad de Santander, 2012
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| dc.title.eng.fl_str_mv |
Two disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa δ-endotoxin increase stability with no effect on toxicity |
| title |
Two disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa δ-endotoxin increase stability with no effect on toxicity |
| spellingShingle |
Two disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa δ-endotoxin increase stability with no effect on toxicity Disulfide Bonds CD Spectra Cry3Aa Site Directed Mutagenesis |
| title_short |
Two disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa δ-endotoxin increase stability with no effect on toxicity |
| title_full |
Two disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa δ-endotoxin increase stability with no effect on toxicity |
| title_fullStr |
Two disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa δ-endotoxin increase stability with no effect on toxicity |
| title_full_unstemmed |
Two disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa δ-endotoxin increase stability with no effect on toxicity |
| title_sort |
Two disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa δ-endotoxin increase stability with no effect on toxicity |
| dc.creator.fl_str_mv |
Wu, Sheng Jiun Florez, Alvaro M. Homoelle, Bradley J. Dean, Donald H. Alzate, Oscar |
| dc.contributor.author.spa.fl_str_mv |
Wu, Sheng Jiun Florez, Alvaro M. Homoelle, Bradley J. Dean, Donald H. Alzate, Oscar |
| dc.subject.proposal.eng.fl_str_mv |
Disulfide Bonds CD Spectra Cry3Aa Site Directed Mutagenesis |
| topic |
Disulfide Bonds CD Spectra Cry3Aa Site Directed Mutagenesis |
| description |
9 p. |
| publishDate |
2012 |
| dc.date.issued.spa.fl_str_mv |
2012-05 |
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2019-08-27T17:32:11Z |
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2019-08-27T17:32:11Z |
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Artículo de revista |
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10.4236/abc.2012.22015. |
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2162-2191 2162-2183 |
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https://repositorio.udes.edu.co/handle/001/3659 |
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10.4236/abc.2012.22015. 2162-2191 2162-2183 |
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https://repositorio.udes.edu.co/handle/001/3659 |
| dc.language.iso.spa.fl_str_mv |
eng |
| language |
eng |
| dc.relation.ispartof.eng.fl_str_mv |
Advances in Biological Chemistry |
| dc.relation.ispartofseries.eng.fl_str_mv |
Advances in Biological Chemistry;N° 2, 123-131, 2012 |
| dc.rights.spa.fl_str_mv |
Derechos Reservados - Universidad de Santander, 2012 |
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Atribución-NoComercial 4.0 Internacional (CC BY-NC 4.0) |
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https://creativecommons.org/licenses/by-nc/4.0/ |
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Derechos Reservados - Universidad de Santander, 2012 Atribución-NoComercial 4.0 Internacional (CC BY-NC 4.0) https://creativecommons.org/licenses/by-nc/4.0/ http://purl.org/coar/access_right/c_abf2 |
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openAccess |
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application/pdf |
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https://www.scirp.org/pdf/ABC20120200004_18831300.pdf |
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Wu, Sheng Jiunf0ca0543-4987-462e-844f-b33023263c58-1Florez, Alvaro M.03b37924-311e-4ca6-bae7-fb60528ecb1c-1Homoelle, Bradley J.fec00867-7717-4b70-ad93-7ce06a3df768-1Dean, Donald H.e9be6363-31b6-435b-8ab0-ad1ca994c335-1Alzate, Oscar73209183-39f5-47d8-b148-c89b5056c6bb-12019-08-27T17:32:11Z2019-08-27T17:32:11Z2012-059 p.To increase protein stability and test protein function, three double-cysteine mutations were individually introduced by protein engineering into the cysteinefree Cry3Aa δ-endotoxin from Bacillus thuringiensis. These mutations were designed to create disulfide bonds between α-helices 2 and 5 (positions 110 - 193), and α-helices 5 and 7 (positions 195 - 276 and 198 - 276). Comparison of the CD spectra of the wild-type and the double-cysteine mutant proteins indicates a tighter helical packing consistent with formation of at least two of the disulfide bonds between the central and the outer helices. Thermal stability analysis indicates that potential covalent linkages between the central α-helix 5 and the other helices increase resistance to thermal denaturation by 10˚C to 14˚C compared to the thermal stability of the wild-type protein. Spectroscopic analysis of the disulfide-specific absorbance band indicates that the double mutant proteins are more stable to temperature and denaturant (guanidine hydrochloride) than the wild-type protein, as a result of the formation of two of the disulfide bridges. These results indicate that the double mutations M110C/F193C and A198C/V276C successfully established disulfide bonds, resulting in a more stable structure of the entire toxin. Despite the increase in stability and structural changes introduced by the disulfide bonds, no effect on toxicity was observed. A possible mechanism involving the insertion of all of domain I of Cry3Aa toxin into the target membrane accounts for these observations.application/pdf10.4236/abc.2012.22015.2162-21912162-2183https://repositorio.udes.edu.co/handle/001/3659engAdvances in Biological ChemistryAdvances in Biological Chemistry;N° 2, 123-131, 2012Derechos Reservados - Universidad de Santander, 2012info:eu-repo/semantics/openAccessAtribución-NoComercial 4.0 Internacional (CC BY-NC 4.0)https://creativecommons.org/licenses/by-nc/4.0/http://purl.org/coar/access_right/c_abf2https://www.scirp.org/pdf/ABC20120200004_18831300.pdfDisulfide BondsCD SpectraCry3AaSite Directed MutagenesisTwo disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa δ-endotoxin increase stability with no effect on toxicityArtículo de revistahttp://purl.org/coar/resource_type/c_6501http://purl.org/coar/resource_type/c_2df8fbb1Textinfo:eu-repo/semantics/articlehttp://purl.org/redcol/resource_type/ARTinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/version/c_970fb48d4fbd8a85PublicationTEXTTwo disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa δ-endotoxin increase stability with no effect on toxicity.pdf.txtTwo disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa δ-endotoxin increase stability with no effect on toxicity.pdf.txtExtracted texttext/plain36481https://repositorio.udes.edu.co/bitstreams/79915aa3-267c-4069-8d59-01322f7a5207/download6c768e8a5119ad823fb48c46bd057ae8MD53THUMBNAILTwo disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa δ-endotoxin increase stability with no effect on toxicity.pdf.jpgTwo disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa δ-endotoxin increase stability with no effect on toxicity.pdf.jpgGenerated Thumbnailimage/jpeg1781https://repositorio.udes.edu.co/bitstreams/a82984e2-9552-465c-bd7e-b4533b62362e/download2e509cda79211bc630bab9ab68e74ea6MD54ORIGINALTwo disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa δ-endotoxin increase stability with no effect on toxicity.pdfTwo disulfide mutants in domain I of Bacillus thuringiensis Cry3Aa δ-endotoxin increase stability with no effect on toxicity.pdfapplication/pdf314406https://repositorio.udes.edu.co/bitstreams/a9506aad-a226-4e41-87f9-0c6a33d568e3/downloadad5ee614436451dd8cb730259a75788cMD55LICENSElicense.txtlicense.txttext/plain; charset=utf-859https://repositorio.udes.edu.co/bitstreams/b0f79685-c9f1-4ff6-ac07-e95371b8efac/download38d94cf55aa1bf2dac1a736ac45c881cMD52001/3659oai:repositorio.udes.edu.co:001/36592023-10-11 11:11:25.366https://creativecommons.org/licenses/by-nc/4.0/Derechos Reservados - Universidad de Santander, 2012https://repositorio.udes.edu.coRepositorio Universidad de Santandersoporte@metabiblioteca.comTGljZW5jaWEgZGUgUHVibGljYWNpw7NuIFVERVMKRGlyZWN0cmljZXMgZGUgVVNPIHkgQUNDRVNPCgo= |