Open AccessLeishmania (Viannia) panamensis expresses a nuclease with molecular and biochemical features similar to the Endonuclease G of higher eukaryotes*
Author(s) -
Miguel Toro-Londoño,
Edwin Patiño,
Sara M. Robledo,
Antonio Jiménez-Ruı́z,
Juan Fernando Alzate
Publication year - 2011
Publication title -
colombia medica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.455
H-Index - 18
eISSN - 1657-9534
pISSN - 0120-8322
DOI - 10.25100/cm.v42i2.766
Subject(s) - biology , endonuclease , nuclease , escherichia coli , gene , leishmania , dna , heterologous expression , recombinant dna , genetics , microbiology and biotechnology , plasmid , biochemistry , parasite hosting , world wide web , computer science
Objective: To characterize the molecular and biochemical features of the Endonuclease G of Leishmania (Viannia) panamensis.Methods: The gene of the putative L. (V.) panamensis Endonuclease G was amplified, cloned, and sequenced. The recombinant protein was produced in a heterologous expression system and biochemical assays were run to determine its ion, temperature, and pH preferences.Results: The L. (V.) panamensis rENDOG has biochemical features similar to those found in other trypanosomatids and higher eukaryotes. In addition, phylogenetic analysis revealed a possible evolutionary relationship with metazoan ENDOG.Conclusions: L. (V.) panamensis has a gene that codifies an ENDOG homologous to those of higher organisms. This enzyme can be produced in Escherichia coli and is able to degrade covalently closed circular double-stranded DNA. It has a magnesium preference, can be inhibited by potassium, and is able to function within a wide temperature and pH range.