Crystallization of the recombinant endonuclease EndoG from Leishmania (Viannia) panamensis
ABSTRACT: Endonuclease G (EndoG) is an enzyme that specifically cleaves double stranded DNA at the dG and dC positions and has been shown to participate in chromatin degradation during apoptosis in Leishmania. The main goal of this work was to purify and crystallize EndoG in preparation for future s...
- Autores:
-
Patiño Márquez, Isabel Andrea
Patiño González, Edwin Bairon
Alzate Restrepo, Juan Fernando
- Tipo de recurso:
- Article of investigation
- Fecha de publicación:
- 2015
- Institución:
- Universidad de Antioquia
- Repositorio:
- Repositorio UdeA
- Idioma:
- eng
- OAI Identifier:
- oai:bibliotecadigital.udea.edu.co:10495/9631
- Acceso en línea:
- http://hdl.handle.net/10495/9631
- Palabra clave:
- Cristalización
Crystallization
Leishmaniasis
EndoG
- Rights
- openAccess
- License
- Atribución-NoComercial-CompartirIgual 2.5 Colombia (CC BY-NC-SA 2.5 CO)
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dc.title.spa.fl_str_mv |
Crystallization of the recombinant endonuclease EndoG from Leishmania (Viannia) panamensis |
dc.title.alternative.spa.fl_str_mv |
Cristalización de la endonucleasa EndoG recombinante de Leishmania (Viannia) panamensis |
title |
Crystallization of the recombinant endonuclease EndoG from Leishmania (Viannia) panamensis |
spellingShingle |
Crystallization of the recombinant endonuclease EndoG from Leishmania (Viannia) panamensis Cristalización Crystallization Leishmaniasis EndoG |
title_short |
Crystallization of the recombinant endonuclease EndoG from Leishmania (Viannia) panamensis |
title_full |
Crystallization of the recombinant endonuclease EndoG from Leishmania (Viannia) panamensis |
title_fullStr |
Crystallization of the recombinant endonuclease EndoG from Leishmania (Viannia) panamensis |
title_full_unstemmed |
Crystallization of the recombinant endonuclease EndoG from Leishmania (Viannia) panamensis |
title_sort |
Crystallization of the recombinant endonuclease EndoG from Leishmania (Viannia) panamensis |
dc.creator.fl_str_mv |
Patiño Márquez, Isabel Andrea Patiño González, Edwin Bairon Alzate Restrepo, Juan Fernando |
dc.contributor.author.none.fl_str_mv |
Patiño Márquez, Isabel Andrea Patiño González, Edwin Bairon Alzate Restrepo, Juan Fernando |
dc.subject.none.fl_str_mv |
Cristalización Crystallization Leishmaniasis EndoG |
topic |
Cristalización Crystallization Leishmaniasis EndoG |
description |
ABSTRACT: Endonuclease G (EndoG) is an enzyme that specifically cleaves double stranded DNA at the dG and dC positions and has been shown to participate in chromatin degradation during apoptosis in Leishmania. The main goal of this work was to purify and crystallize EndoG in preparation for future structural studies that will permit a detailed understanding of the function of this enzyme. Materials and methods: EndoG protein was puri ed using Ni-af nity chromatography under denaturing conditions, then refolded in vitro and crystallized by the hanging-drop vapor diffusion method. Results and conclusion: The endonuclease G protein from Leishmania (viannia) panamensis was overexpressed, refolded, purified and demonstrated to be enzymatically active. Here, we reports the first successful crystallization of the EndoG protein in this group of protozoan parasites. The protein was crystallized by the hanging-drop vapor diffusion method. High quality EndoG crystals were obtained that perhaps will permit determination of the three-dimensional structure of EndoG using X-ray diffraction. |
publishDate |
2015 |
dc.date.issued.none.fl_str_mv |
2015 |
dc.date.accessioned.none.fl_str_mv |
2018-06-20T15:16:59Z |
dc.date.available.none.fl_str_mv |
2018-06-20T15:16:59Z |
dc.type.spa.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.coarversion.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
dc.type.hasversion.spa.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.coar.spa.fl_str_mv |
http://purl.org/coar/resource_type/c_2df8fbb1 |
dc.type.redcol.spa.fl_str_mv |
https://purl.org/redcol/resource_type/ART |
dc.type.local.spa.fl_str_mv |
Artículo de investigación |
format |
http://purl.org/coar/resource_type/c_2df8fbb1 |
status_str |
publishedVersion |
dc.identifier.citation.spa.fl_str_mv |
Patiño I, Patiño E, Alzate JF. Crystallization of the recombinant endonuclease EndoG from Leishmania (Viannia) panamensis. Actual. Biol; 2015; 37 (102): 27-32. |
dc.identifier.issn.none.fl_str_mv |
0304-3584 |
dc.identifier.uri.none.fl_str_mv |
http://hdl.handle.net/10495/9631 |
dc.identifier.eissn.none.fl_str_mv |
2145-7166 |
identifier_str_mv |
Patiño I, Patiño E, Alzate JF. Crystallization of the recombinant endonuclease EndoG from Leishmania (Viannia) panamensis. Actual. Biol; 2015; 37 (102): 27-32. 0304-3584 2145-7166 |
url |
http://hdl.handle.net/10495/9631 |
dc.language.iso.spa.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartofjournalabbrev.spa.fl_str_mv |
Actual. Biol. |
dc.rights.*.fl_str_mv |
Atribución-NoComercial-CompartirIgual 2.5 Colombia (CC BY-NC-SA 2.5 CO) |
dc.rights.spa.fl_str_mv |
info:eu-repo/semantics/openAccess |
dc.rights.uri.*.fl_str_mv |
https://creativecommons.org/licenses/by-nc-sa/2.5/co/ |
dc.rights.accessrights.spa.fl_str_mv |
http://purl.org/coar/access_right/c_abf2 |
dc.rights.creativecommons.spa.fl_str_mv |
https://creativecommons.org/licenses/by-nc-sa/4.0/ |
rights_invalid_str_mv |
Atribución-NoComercial-CompartirIgual 2.5 Colombia (CC BY-NC-SA 2.5 CO) https://creativecommons.org/licenses/by-nc-sa/2.5/co/ http://purl.org/coar/access_right/c_abf2 https://creativecommons.org/licenses/by-nc-sa/4.0/ |
eu_rights_str_mv |
openAccess |
dc.format.mimetype.spa.fl_str_mv |
application/pdf |
dc.publisher.spa.fl_str_mv |
Universidad de Antioquia, Facultad de Ciencias Exactas y Naturales, Instituto de Biología |
dc.publisher.group.spa.fl_str_mv |
Grupo Interdisciplinario de Estudios Moleculares Grupo de Bioquímica Estructural de Macromoléculas |
dc.publisher.place.spa.fl_str_mv |
Medellín, Colombia |
institution |
Universidad de Antioquia |
bitstream.url.fl_str_mv |
http://bibliotecadigital.udea.edu.co/bitstream/10495/9631/1/PatinnoMarquezIsabel_2015_CrystallizationEndonucleaseEndoG.pdf http://bibliotecadigital.udea.edu.co/bitstream/10495/9631/2/license_url http://bibliotecadigital.udea.edu.co/bitstream/10495/9631/3/license_text http://bibliotecadigital.udea.edu.co/bitstream/10495/9631/4/license_rdf http://bibliotecadigital.udea.edu.co/bitstream/10495/9631/5/license.txt |
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andres.perez@udea.edu.co |
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spelling |
Patiño Márquez, Isabel AndreaPatiño González, Edwin BaironAlzate Restrepo, Juan Fernando2018-06-20T15:16:59Z2018-06-20T15:16:59Z2015Patiño I, Patiño E, Alzate JF. Crystallization of the recombinant endonuclease EndoG from Leishmania (Viannia) panamensis. Actual. Biol; 2015; 37 (102): 27-32.0304-3584http://hdl.handle.net/10495/96312145-7166ABSTRACT: Endonuclease G (EndoG) is an enzyme that specifically cleaves double stranded DNA at the dG and dC positions and has been shown to participate in chromatin degradation during apoptosis in Leishmania. The main goal of this work was to purify and crystallize EndoG in preparation for future structural studies that will permit a detailed understanding of the function of this enzyme. Materials and methods: EndoG protein was puri ed using Ni-af nity chromatography under denaturing conditions, then refolded in vitro and crystallized by the hanging-drop vapor diffusion method. Results and conclusion: The endonuclease G protein from Leishmania (viannia) panamensis was overexpressed, refolded, purified and demonstrated to be enzymatically active. Here, we reports the first successful crystallization of the EndoG protein in this group of protozoan parasites. The protein was crystallized by the hanging-drop vapor diffusion method. High quality EndoG crystals were obtained that perhaps will permit determination of the three-dimensional structure of EndoG using X-ray diffraction.RESUMEN: Antecedentes y objetivos: La endonucleasa G (EndoG) es una enzima que escinde específicamente en las posiciones dG y dC del ADN de cadena doble y se ha demostrado que participa en la degradación de la cromatina durante el proceso de apoptosis en Leishmania. El objetivo principal de este trabajo fue la purificación y cristalización de EndoG como preámbulo para los estudios estructurales futuros que permitan entender detalladamente el funcionamiento de esta enzima. Materiales y métodos: La proteína EndoG fue purificada en condiciones desnaturalizantes usando cromatografía de Ni, luego fue renaturalizada in vitro y cristalizada por el método de difusión de vapor por gota colgante. Resultados y conclusión: La proteína EndoG de Leishmania (viannia) panamensis fue sobreexpresada, renaturalizada, purificada y demostró estar enzimáticamente activa. Aquí, se registra la primera cristalización exitosa de la proteína EndoG de este grupo de parásitos protozoarios. La proteína fue cristalizada por el método de difusión de vapor por gota colgante. Se obtuvieron cristales de alta calidad de EndoG que posiblemente nos permitirán determinar la estructura tridimensional de EndoG usando difracción de rayos-X.COL0156275COL0007462application/pdfengUniversidad de Antioquia, Facultad de Ciencias Exactas y Naturales, Instituto de BiologíaGrupo Interdisciplinario de Estudios MolecularesGrupo de Bioquímica Estructural de MacromoléculasMedellín, Colombiainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_2df8fbb1https://purl.org/redcol/resource_type/ARTArtículo de investigaciónhttp://purl.org/coar/version/c_970fb48d4fbd8a85Atribución-NoComercial-CompartirIgual 2.5 Colombia (CC BY-NC-SA 2.5 CO)info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/co/http://purl.org/coar/access_right/c_abf2https://creativecommons.org/licenses/by-nc-sa/4.0/CristalizaciónCrystallizationLeishmaniasisEndoGCrystallization of the recombinant endonuclease EndoG from Leishmania (Viannia) panamensisCristalización de la endonucleasa EndoG recombinante de Leishmania (Viannia) panamensisActual. Biol.Actualidades Biológicas273237102ORIGINALPatinnoMarquezIsabel_2015_CrystallizationEndonucleaseEndoG.pdfPatinnoMarquezIsabel_2015_CrystallizationEndonucleaseEndoG.pdfArtículo de investigaciónapplication/pdf2091093http://bibliotecadigital.udea.edu.co/bitstream/10495/9631/1/PatinnoMarquezIsabel_2015_CrystallizationEndonucleaseEndoG.pdf5ce098652fdd65ec177e78c78b5c9c83MD51CC-LICENSElicense_urllicense_urltext/plain; charset=utf-849http://bibliotecadigital.udea.edu.co/bitstream/10495/9631/2/license_url4afdbb8c545fd630ea7db775da747b2fMD52license_textlicense_texttext/html; charset=utf-80http://bibliotecadigital.udea.edu.co/bitstream/10495/9631/3/license_textd41d8cd98f00b204e9800998ecf8427eMD53license_rdflicense_rdfLicenciaapplication/rdf+xml; charset=utf-80http://bibliotecadigital.udea.edu.co/bitstream/10495/9631/4/license_rdfd41d8cd98f00b204e9800998ecf8427eMD54LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://bibliotecadigital.udea.edu.co/bitstream/10495/9631/5/license.txt8a4605be74aa9ea9d79846c1fba20a33MD5510495/9631oai:bibliotecadigital.udea.edu.co:10495/96312021-07-16 22:56:28.829Repositorio Institucional Universidad de Antioquiaandres.perez@udea.edu.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 |