Preferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane Vesicles
13 p.
- Autores:
-
Hussain, Syed-Rehan A.
Florez, Alvaro M.
Dean, Donald H.
Alzate, Oscar
- Tipo de recurso:
- Article of journal
- Fecha de publicación:
- 2010
- Institución:
- Universidad de Santander
- Repositorio:
- Repositorio Universidad de Santander
- Idioma:
- eng
- OAI Identifier:
- oai:repositorio.udes.edu.co:001/3728
- Acceso en línea:
- https://repositorio.udes.edu.co/handle/001/3728
- Palabra clave:
- Bacillus thuringiensis
Site directed mutagenesis
δ -endotoxin
Mutagénesis sitio dirigida
- Rights
- openAccess
- License
- Derechos Reservados - Revista Colombiana de Biotecnología, 2010
| id |
RUDES2_bb29deb694250d27fc42f2722da16b9b |
|---|---|
| oai_identifier_str |
oai:repositorio.udes.edu.co:001/3728 |
| network_acronym_str |
RUDES2 |
| network_name_str |
Repositorio Universidad de Santander |
| repository_id_str |
|
| dc.title.eng.fl_str_mv |
Preferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane Vesicles |
| dc.title.alternative.spa.fl_str_mv |
Protección preferencial de los dominios II y III de la toxina Cry1Aa de Bacillus thuringiensisen Vesículas de Membrana de Borde de Cepillo |
| title |
Preferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane Vesicles |
| spellingShingle |
Preferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane Vesicles Bacillus thuringiensis Site directed mutagenesis δ -endotoxin Mutagénesis sitio dirigida |
| title_short |
Preferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane Vesicles |
| title_full |
Preferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane Vesicles |
| title_fullStr |
Preferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane Vesicles |
| title_full_unstemmed |
Preferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane Vesicles |
| title_sort |
Preferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane Vesicles |
| dc.creator.fl_str_mv |
Hussain, Syed-Rehan A. Florez, Alvaro M. Dean, Donald H. Alzate, Oscar |
| dc.contributor.author.spa.fl_str_mv |
Hussain, Syed-Rehan A. Florez, Alvaro M. Dean, Donald H. Alzate, Oscar |
| dc.subject.proposal.eng.fl_str_mv |
Bacillus thuringiensis Site directed mutagenesis δ -endotoxin |
| topic |
Bacillus thuringiensis Site directed mutagenesis δ -endotoxin Mutagénesis sitio dirigida |
| dc.subject.proposal.spa.fl_str_mv |
Mutagénesis sitio dirigida |
| description |
13 p. |
| publishDate |
2010 |
| dc.date.issued.spa.fl_str_mv |
2010-12 |
| dc.date.accessioned.spa.fl_str_mv |
2019-09-03T15:28:24Z |
| dc.date.available.spa.fl_str_mv |
2019-09-03T15:28:24Z |
| dc.type.spa.fl_str_mv |
Artículo de revista |
| dc.type.coar.fl_str_mv |
http://purl.org/coar/resource_type/c_2df8fbb1 |
| dc.type.coarversion.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
| dc.type.coar.spa.fl_str_mv |
http://purl.org/coar/resource_type/c_6501 |
| dc.type.content.spa.fl_str_mv |
Text |
| dc.type.driver.spa.fl_str_mv |
info:eu-repo/semantics/article |
| dc.type.redcol.spa.fl_str_mv |
http://purl.org/redcol/resource_type/ART |
| dc.type.version.spa.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
| format |
http://purl.org/coar/resource_type/c_6501 |
| status_str |
publishedVersion |
| dc.identifier.issn.spa.fl_str_mv |
1909-8758 0123-3475 |
| dc.identifier.uri.spa.fl_str_mv |
https://repositorio.udes.edu.co/handle/001/3728 |
| identifier_str_mv |
1909-8758 0123-3475 |
| url |
https://repositorio.udes.edu.co/handle/001/3728 |
| dc.language.iso.spa.fl_str_mv |
eng |
| language |
eng |
| dc.relation.ispartof.spa.fl_str_mv |
Revista Colombiana de Biotecnología |
| dc.relation.ispartofseries.spa.fl_str_mv |
Revista Colombiana de Biotecnología;Volumen 12, Número 2, p. 14-26, 2010 |
| dc.rights.spa.fl_str_mv |
Derechos Reservados - Revista Colombiana de Biotecnología, 2010 |
| dc.rights.coar.fl_str_mv |
http://purl.org/coar/access_right/c_abf2 |
| dc.rights.accessrights.spa.fl_str_mv |
info:eu-repo/semantics/openAccess |
| dc.rights.creativecommons.spa.fl_str_mv |
Atribución-NoComercial 4.0 Internacional (CC BY-NC 4.0) |
| dc.rights.uri.spa.fl_str_mv |
https://creativecommons.org/licenses/by-nc/4.0/ |
| rights_invalid_str_mv |
Derechos Reservados - Revista Colombiana de Biotecnología, 2010 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 |
| eu_rights_str_mv |
openAccess |
| dc.format.mimetype.spa.fl_str_mv |
application/pdf |
| dc.source.eng.fl_str_mv |
https://revistas.unal.edu.co/index.php/biotecnologia/article/view/18424/19938 |
| institution |
Universidad de Santander |
| bitstream.url.fl_str_mv |
https://repositorio.udes.edu.co/bitstreams/d0edc602-6b6d-4cf5-a1fa-73361b8784e3/download https://repositorio.udes.edu.co/bitstreams/402bfde0-cacd-4e16-9dc2-fd118b2f5b83/download https://repositorio.udes.edu.co/bitstreams/d7ab16dd-59c7-4ffb-99a4-73d8118dd441/download https://repositorio.udes.edu.co/bitstreams/769b4010-3e5e-4272-aeff-44754f8ecdd3/download |
| bitstream.checksum.fl_str_mv |
2ea160f621487f655473a948b7bf094a 51b370554cca4a983c391a5867fbe26a 8442fd9fb877b4c6d1180041934665a7 38d94cf55aa1bf2dac1a736ac45c881c |
| bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 |
| repository.name.fl_str_mv |
Repositorio Universidad de Santander |
| repository.mail.fl_str_mv |
soporte@metabiblioteca.com |
| _version_ |
1814158550748889088 |
| spelling |
Hussain, Syed-Rehan A.af346681-389f-42aa-ada3-bd368aa0c9d1-1Florez, Alvaro M.03b37924-311e-4ca6-bae7-fb60528ecb1c-1Dean, Donald H.e9be6363-31b6-435b-8ab0-ad1ca994c335-1Alzate, Oscar73209183-39f5-47d8-b148-c89b5056c6bb-12019-09-03T15:28:24Z2019-09-03T15:28:24Z2010-1213 p.The surface exposed Leucine 371 on loop 2 of domain II, in Cry1Aa toxin, was mutated to Lysine to generate the trypsin-sensitive mutant, L371K. Upon trypsin digestion L371K is cleaved into approxima-tely 37 and 26 kDa fragments. These are separable on SDS-PAGE, but remain as a single molecule of 65 kDa upon purification by liquid chromatography. The larger fragment is domain I and a portion of domain II (amino acid residues 1 to 371). The smaller 26-kDa polypeptide is the remainder of domain II and domain III (amino acids 372 to 609). When the mutant toxin was treated with high dose of M. sexta gut juice both fragments were degraded. However, when incubated with M. sexta BBMV, the 26 kDa fragment (domains II and III) was preferentially protected from gut juice proteases. As previously reported, wild type Cry1Aa toxin was also protected against degradation by gut juice proteases when incubated with M. sexta BBMV. On the contrary, when mouse BBMV was added to the reaction mixture neither Cry1Aa nor L371K toxins showed resistance to M. sexta gut juice proteases and were degraded. Since the whole Cry1Aa toxin and most of the domain II and domain III of L371K are protected from proteases in the presence of BBMV of the target insect, we suggest that the insertion of the toxin into the membrane is complex and involves all three domains.La superficie de la toxina Cry1Aa, en el asa 2 del dominio II contiene expuesta la leucina 371, la cual fue mo-dificada a lisina produciendo una mutante sensible a la tripsina, L371K. Esta mutante produce dos fragmen-tos de 37 y 26 kDa por acción de la tripsina que son separables por SDS-PAGE, pero que a la purificación por cromatografía líquida se mantienen como una sola molécula de 65 kDa. El fragmento grande contiene al dominio I y una parte del dominio II (aminoácidos 1 al 371). El polipéptido de 26 kDa contiene la parte res-tante del dominio II y dominio III (aminoácidos 372 al 609). Cuando la toxina mutante fue tratada con dosis altas de jugo intestinal de Manduca sexta, ambos fragmentos fueron degradados. Sin embargo, cuando fueron incubados en VMBC de M. sexta, el fragmento de 26 kDa fue protegido preferencialmente de las proteasas intestinales. Como se ha reportado, la toxina silvestre Cry1Aa también es protegida de la degradación de las proteasas cuando es incubada en VMBC de M. sexta. Sin embargo, cuando se adicionó VMBC de ratón a la mezcla de reacción, ni la toxina Cry1Aa ni la mutante L371K mostraron resistencia a las proteasas y fueron degradadas. Dado que la toxina completa de Cry1Aa y casi todo de los dominios II y III de L371K están protegidos de proteasas en presencia de VMBC del insecto, este estudio sugiere que la inserción de la toxina en la membrana involucra los tres dominios.application/pdf1909-87580123-3475https://repositorio.udes.edu.co/handle/001/3728engRevista Colombiana de BiotecnologíaRevista Colombiana de Biotecnología;Volumen 12, Número 2, p. 14-26, 2010Derechos Reservados - Revista Colombiana de Biotecnología, 2010info: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://revistas.unal.edu.co/index.php/biotecnologia/article/view/18424/19938Bacillus thuringiensisSite directed mutagenesisδ -endotoxinMutagénesis sitio dirigidaPreferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane VesiclesProtección preferencial de los dominios II y III de la toxina Cry1Aa de Bacillus thuringiensisen Vesículas de Membrana de Borde de CepilloArtículo 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_970fb48d4fbd8a85PublicationTEXTPreferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane Vesicles.pdf.txtPreferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane Vesicles.pdf.txtExtracted texttext/plain39374https://repositorio.udes.edu.co/bitstreams/d0edc602-6b6d-4cf5-a1fa-73361b8784e3/download2ea160f621487f655473a948b7bf094aMD53THUMBNAILPreferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane Vesicles.pdf.jpgPreferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane Vesicles.pdf.jpgGenerated Thumbnailimage/jpeg1498https://repositorio.udes.edu.co/bitstreams/402bfde0-cacd-4e16-9dc2-fd118b2f5b83/download51b370554cca4a983c391a5867fbe26aMD54ORIGINALPreferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane Vesicles.pdfPreferential Protection of Domains II and III of Bacillus thuringiensis Cry1Aa Toxin by Brush Border Membrane Vesicles.pdfapplication/pdf284014https://repositorio.udes.edu.co/bitstreams/d7ab16dd-59c7-4ffb-99a4-73d8118dd441/download8442fd9fb877b4c6d1180041934665a7MD55LICENSElicense.txtlicense.txttext/plain; charset=utf-859https://repositorio.udes.edu.co/bitstreams/769b4010-3e5e-4272-aeff-44754f8ecdd3/download38d94cf55aa1bf2dac1a736ac45c881cMD52001/3728oai:repositorio.udes.edu.co:001/37282023-10-09 10:23:43.689https://creativecommons.org/licenses/by-nc/4.0/Derechos Reservados - Revista Colombiana de Biotecnología, 2010https://repositorio.udes.edu.coRepositorio Universidad de Santandersoporte@metabiblioteca.comTGljZW5jaWEgZGUgUHVibGljYWNpw7NuIFVERVMKRGlyZWN0cmljZXMgZGUgVVNPIHkgQUNDRVNPCgo= |