Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the mitochondrial tryparedoxin peroxidase from Leishmania braziliensis.

Acta Crystallogr Sect F Struct Biol Cryst Commun

Laboratório Nacional de Biociências, Laboratório Nacional de Luz Síncrotron, Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP 13083-970, Brazil.

Published: April 2013


Category Ranking

98%

Total Visits

921

Avg Visit Duration

2 minutes

Citations

20

Article Abstract

Tryparedoxin peroxidase (TXNPx) is an essential constituent of the main enzymatic scavenger system for reactive oxygen species (ROS) in trypanosomatids. Genetic studies have demonstrated the importance of this system for the development and virulence of these parasites, representing a potential target for the discovery of new trypanocidal drugs. In this work, the mitochondrial TXNPx from Leishmania braziliensis was cloned, overexpressed, purified and crystallized. The crystals diffracted to 3.3 Å resolution and belonged to space group P4(2)2(1)2, with unit-cell parameters a = b = 131.8, c = 44.4 Å. These studies will contribute to a better understanding of the molecular mechanisms involved in ROS detoxification by trypanosomatids.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3614166PMC
http://dx.doi.org/10.1107/S1744309113003989DOI Listing

Publication Analysis

Top Keywords

tryparedoxin peroxidase
8
leishmania braziliensis
8
cloning expression
4
expression purification
4
purification crystallization
4
crystallization preliminary
4
preliminary x-ray
4
x-ray diffraction
4
diffraction analysis
4
analysis mitochondrial
4

Similar Publications

Background: Leishmaniasis continues to pose a significant global health challenge, exacerbated by the increasing resistance to current therapeutic agents such as miltefosine and amphotericin B. This growing resistance highlights the urgent need for alternative treatment strategies. In this context, phytomedicine has emerged as a promising avenue for novel antileishmanial therapies.

View Article and Find Full Text PDF

Neglected tropical diseases such as Leishmaniasis and Chagas disease remain critical public health challenges. This study applied ligand-based virtual screening to a dataset of 4,150 secondary metabolites from the Lamiaceae family, aiming to identify multitarget molecules against four Leishmania species (L. infantum, L.

View Article and Find Full Text PDF

Background: Increasing resistance to antimonial drugs has become a significant challenge in effective treatment in endemic regions of leishmaniasis. Antioxidant defense plays a crucial role in antimony resistance by combating antimonial-induced oxidative stress. Accordingly, we investigated the transcript levels of some antioxidant genes in antimony-sensitive and resistant clinical Leishmania tropica isolates.

View Article and Find Full Text PDF

Molecular and biochemical approaches of the trypanothione system in Leishmania spp.: A key player in parasite resistance to antimonial therapy.

Biochimie

July 2025

Fundação Oswaldo Cruz, Instituto Oswaldo Cruz, Laboratório de Biologia Molecular e Doenças Endêmicas, Rio de Janeiro, RJ, Brazil. Electronic address:

The trypanothione system is a crucial antioxidant defense mechanism in Leishmania spp. The enzymes involved, including trypanothione reductase (TR), trypanothione synthetase (TS), tryparedoxin (TXN) and tryparedoxin peroxidase (TXNPx) are essential for maintaining the redox balance. This system plays a fundamental role in the biology of Leishmania spp.

View Article and Find Full Text PDF

We employed a CRISPR/Cas9 technique in Leishmania major to evaluate its efficiency in editing a kDNA-associated gene, the universal minicircle sequence binding protein (UMSBP), which is involved in mitochondrial respiration and kinetoplast division. Using this toolkit, we generated UMSBP mNG-tagged and single knockout L. major (LmUMSBP) parasites, which were confirmed by PCR, confocal microscopy and Western blot analyses.

View Article and Find Full Text PDF