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...

Full description

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)
Description
Summary: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.