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LdIscU is a [2Fe-2S] scaffold protein which interacts with LdIscS and its expression is modulated by Fe-S proteins in

Krishn Pratap Singh¹, Shadab Anwar², Amir Zaidi¹

¹Laboratory of Molecular Biochemistry and Cell Biology, Department of Biochemistry, Rajendra Memorial Research Institute of Medical Sciences (I.C.M.R.), Agamkuan, Patna 800007, India.

International Journal of Biological Macromolecules

|May 22, 2018

Abstract:

The pathogenicity of protozoan parasites is frequently attributed to their ability to circumvent the deleterious effects of ROS and Fe-S clusters are among their susceptible targets with paramount importance for parasite survival. The biogenesis of Fe-S clusters is orchestrated by ISC system; the sulfur donor IscS and scaffold protein IscU being its core components. However, among protozoan parasites including Leishmania, no information is available regarding biochemical aspect of IscU, its interaction partners and regulation. Here, we show that Leishmania donovani IscU homolog, LdIscU, readily assembles [2Fe-2S] clusters and, interestingly, follows Michaelis-Menten enzyme kinetics. It is localized in the mitochondria of the parasite and interacts with LdIscS to form a stable complex. Additionally, LdIscU and Fe-S proteins activity is significantly upregulated in resistant isolates and during stationary growth stage indicating an association between them. The differential expression of LdIscU modulated by Fe-S proteins demand suggests its potential role in parasite survival and drug resistance. Thus, our study provides novel insight into the Fe-S scaffold protein of a protozoan parasite.

Insights

Leishmania donovani IscU protein assembles iron-sulfur (Fe-S) clusters and is crucial for parasite survival. Its upregulation in drug-resistant parasites suggests a role in drug resistance.

Area of Science:

Biochemistry
Parasitology
Molecular Biology

Background:

Protozoan parasites cause disease by evading reactive oxygen species (ROS), with iron-sulfur (Fe-S) clusters being vital for their survival.
The biogenesis of Fe-S clusters relies on the ISC system, involving IscS (sulfur donor) and IscU (scaffold protein).
Biochemical data on IscU, its interactions, and regulation in protozoan parasites like Leishmania are lacking.

Purpose of the Study:

To investigate the biochemical properties and function of the Leishmania donovani IscU homolog (LdIscU).
To explore the interaction partners and regulatory mechanisms of LdIscU.
To determine the role of LdIscU in parasite survival and drug resistance.

Main Methods:

Characterization of LdIscU's ability to assemble [2Fe-2S] clusters.
Enzyme kinetics analysis of LdIscU using Michaelis-Menten kinetics.
Subcellular localization studies of LdIscU within the parasite.
Co-immunoprecipitation assays to identify LdIscU interaction partners, specifically LdIscS.
Analysis of LdIscU and Fe-S proteins activity in drug-resistant versus sensitive parasite isolates and during different growth stages.

Main Results:

LdIscU successfully assembles [2Fe-2S] clusters and exhibits Michaelis-Menten enzyme kinetics.
LdIscU is localized in the mitochondria and forms a stable complex with LdIscS.
Activity of LdIscU and Fe-S proteins is significantly upregulated in drug-resistant Leishmania isolates and during the stationary growth phase.
Differential expression of LdIscU correlates with Fe-S protein demand.

Conclusions:

LdIscU is a functional Fe-S scaffold protein in Leishmania donovani, essential for parasite survival.
LdIscU interacts with LdIscS, forming a key complex within the parasite's mitochondria.
The upregulation of LdIscU in resistant strains suggests its involvement in Leishmania drug resistance mechanisms.

Read full analysis

Keywords:

Fe-S scaffold protein Iron sulfur clusters Leishmania

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