Overexpression in Escherichia coli and purification of pteridine reductase (PTR1) from a clinical isolate of Leishmania donovani.

Pranav Kumar, Hema Kothari, Neeloo Singh

Drug Target Discovery and Development Division, Central Drug Research Institute, Lucknow, India.

Journal Article: Protein Expression and Purification (impact factor: 1.56). 01/2005; 38(2):228-36. DOI: 10.1016/j.pep.2004.08.015

Abstract

Pteridine reductase 1 (PTR1) is part of a novel metabolic pathway in Leishmania associated with folate metabolism. Its main function is to salvage pterins but a second one is to reduce folates. The novelty and possible uniqueness of the pathway in which PTR1 is involved opens the possibility of developing specific inhibitors, which in combination with dihydrofolate reductase inhibitors could be highly effective against Leishmania. In order to increase our understanding of this putative important chemotherapeutic target, we present here the cloning, overexpression and purification of this enzyme from a clinical isolate of Leishmania donovani causing kala azar in India. This recombinant enzyme will set the basis for inhibition studies as well as for structure-function relationships.

Source: PubMed

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Keywords

chemotherapeutic target
 
cloning
 
folate metabolism
 
inhibition studies
 
kala azar
 
Leishmania
 
Leishmania donovani
 
novel metabolic pathway
 
novelty
 
pathway
 
Pteridine reductase 1
 
PTR1
 
purification
 
recombinant enzyme
 
specific inhibitors
 
structure-function relationships