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...

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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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oai_identifier_str oai:bibliotecadigital.udea.edu.co:10495/9631
network_acronym_str UDEA2
network_name_str Repositorio UdeA
repository_id_str
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
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dc.type.local.spa.fl_str_mv Artículo de investigación
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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
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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/
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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
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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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